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<p>目的のタンパク質のみをターゲットとすることが重要であるため、CUT＆Tagアッセイの成功には非常に特異的な抗体が不可欠です。 厳密な検証基準を満たしている特異性が検証されているDiagenode ChIP-seqグレードの抗体は、CUT＆Tagアッセイでの高いパフォーマンスを保証します。</p>
<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p>目的のタンパク質のみをターゲットとすることが重要であるため、CUT＆Tagアッセイの成功には非常に特異的な抗体が不可欠です。 厳密な検証基準を満たしている特異性が検証されているDiagenode ChIP-seqグレードの抗体は、CUT＆Tagアッセイでの高いパフォーマンスを保証します。</p>
<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For flexibility, the fusion protein is not pre-loaded with sequencing adapters. Please follow the recommended <a href="https://www.diagenode.com/en/documents/assembly-protocol-patn5-20042020-rms" target="_blank">assembly protocol</a> prior to use in CUT&Tag or similar assays (ACT-seq, CoBATCH, TIP-seq and other).</p>
<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> <span>- compatible with histome marks and some transcription factors</span></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-a.jpg" width="600" height="265" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">B.<br /><center><img alt="pA-Tn5 Transposase unloaded" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-b.jpg" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> - compatible with histome marks and some transcription factors</li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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'description' => '<p><span>The negative Ctrl IgG from rabbit has been extensively validated in chromatin immunoprecipitation assays (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy animals. This IgG preparation is intended for use as a negative control in ChIP experiments for specific antibodies made in rabbit. The negative Ctrl IgG from rabbit should be used for ChIP in parallel with specific antibody at the same concentration as the specific antibody. </span></p>
<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-chip.jpg" alt="Rabbit IgG Antibody ChIP Grade" /></p>
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<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'info2' => '<p>The negative control IgG from rabbit has been extensively validated in chromatin immunoprecipitation (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy rabbits. This IgG preparation is intended for use as a negative control in ChIP, MeDIP, IF and other experiments performed with specific antibodies made in rabbit. The negative control IgG from rabbit should be used in parallel with the specific antibody at the same concentration. It is also included in many of our ChIP and MeDIP kits.</p>',
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'name' => '24 UDI for Tagmented libraries - Set I',
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<p>The <strong>24 UDI for tagmented libraries</strong> includes 24 primer pairs for unique dual-indexing allowing the multiplexing of up to <b>24 samples </b>for sequencing on Illumina platforms. These UDI are designed and validated to be used with <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">μChIPmentation for Histones</a> (Cat. No. C01011011), <a href="https://www.diagenode.com/en/p/chipmentation-kit-for-histones">ChIPmentation Kit for Histones</a> (Cat. No. C01011009), <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a> (Cat. No. C01011030), <a href="https://www.diagenode.com/en/categories/atac-seq">ATAC-seq Kit</a> (Cat. No. C01080002). The 24 UDI for tagmented libraries are compatible with other <b>tagmentation</b><b>-based library preparation </b>protocols, such as <a href="https://www.diagenode.com/en/categories/cutandtag">CUT&Tag</a> technologies.</p>
<p>3 sets of UDI for tagmented libraries are available:</p>
<p><strong>24 UDI for tagmented libraries - Set I</strong><br /> <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for tagmented libraries - Set II</a><br /><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3" target="_blank">24 UDI for tagmented libraries - Set III</a><br /><br /></p>
<p><span>Each set can be used for library multiplexing up to 24. All sets can be used simultaneously for library multiplexing up to 72.</span></p>
<p>Features:</p>
<ul>
<li>Multiplexing: <b>up to 72 samples </b>(using all 3 sets simultaneously)<b><br /></b></li>
<li>Allow for <b>identification of index hopping</b></li>
<li>Compatibility: <b>tagmentation</b><b>-based library preparation protocols</b></li>
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'info1' => '<p>The <b>24 UDI (Unique dual indexes) for </b><b>tagmented</b><b> libraries – Set I </b>is compatible with any <b>tagmentation</b><b>-based library preparation </b>protocols, such as <strong>ChIPmentation</strong>, <b>ATAC-seq</b> or <b>CUT&Tag</b> technologies.</p>
<p>The <b>24 UDI for </b><b>tagmented</b><b> libraries </b>provides combinations of barcodes where each barcode is uniquely attributed to one sample. This is a great tool to identify mistakes during index sequencing. A phenomenon, known as index hopping, can lead to misattribution of some reads to the wrong sample. This is particularly frequent with the NovaSeq6000, and thus the use of Unique Dual Indexing (UDI) is highly recommended when using this sequencer.</p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/UDI-for-tagmented-fig1.png" /></center>
<p></p>
<p><small><strong>Figure 1. Sequencing profiles of µChIPmentation libraries generated with 24 UDI for Tagmented libraries</strong> Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 UDI for Tagmented libraries – Set I (Cat. No. Cat. No. C01011034) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003) and rabbit IgG (Cat. No. C15410206) have been used. </small></p>
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'name' => 'Tagmentase (Tn5 transposase) - unloaded',
'description' => '<div class="extra-spaced"><center><img alt="Tagmentase (Tn5 transposase)" src="https://www.diagenode.com/img/banners/banner-tagmentase.jpg" caption="false" width="787" height="236" /></center></div>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
<p><a href="https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf" target="_blank">Protocol for transposome assembly</a></p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
</ul>
<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><img alt="Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1a.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="653" height="282" /></p>
<p><img alt="Tagmentase Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1b.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="645" height="278" /></p>
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<div class="small-12 medium-12 large-12 columns"><center><img alt="Tn5 transposase perfect for NGS" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure2.jpg" width="754" height="492" /></center></div>
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<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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'info2' => '<p><strong>Tagmentase (Tn5 transposase) - unloaded</strong></p>
<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
<p><strong>Storage conditions:</strong> Store at -20°C. Guaranteed stable for 6 months from date of receipt when stored properly.</p>
<p><strong>Storage buffer: </strong>Supplied in solution containing 50% v/v glycerol.</p>
<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones (developped for histone marks and some non-histone proteins), but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<br /> <a href="https://www.diagenode.com/files/application_notes/AN-iDealCUTandTag.pdf"><img src="https://www.diagenode.com/img/banners/cutandtag-appnote.png" /></a><br /><br /></div>
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<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
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<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
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<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
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<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
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<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
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<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
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<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p>目的のタンパク質のみをターゲットとすることが重要であるため、CUT＆Tagアッセイの成功には非常に特異的な抗体が不可欠です。 厳密な検証基準を満たしている特異性が検証されているDiagenode ChIP-seqグレードの抗体は、CUT＆Tagアッセイでの高いパフォーマンスを保証します。</p>
<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> <span>- compatible with histome marks and some transcription factors</span></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-a.jpg" width="600" height="265" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">B.<br /><center><img alt="pA-Tn5 Transposase unloaded" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-b.jpg" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> - compatible with histome marks and some transcription factors</li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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'description' => '<p><span>The negative Ctrl IgG from rabbit has been extensively validated in chromatin immunoprecipitation assays (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy animals. This IgG preparation is intended for use as a negative control in ChIP experiments for specific antibodies made in rabbit. The negative Ctrl IgG from rabbit should be used for ChIP in parallel with specific antibody at the same concentration as the specific antibody. </span></p>
<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-chip.jpg" alt="Rabbit IgG Antibody ChIP Grade" /></p>
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<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'info2' => '<p>The negative control IgG from rabbit has been extensively validated in chromatin immunoprecipitation (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy rabbits. This IgG preparation is intended for use as a negative control in ChIP, MeDIP, IF and other experiments performed with specific antibodies made in rabbit. The negative control IgG from rabbit should be used in parallel with the specific antibody at the same concentration. It is also included in many of our ChIP and MeDIP kits.</p>',
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'name' => '24 UDI for Tagmented libraries - Set I',
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<p>The <strong>24 UDI for tagmented libraries</strong> includes 24 primer pairs for unique dual-indexing allowing the multiplexing of up to <b>24 samples </b>for sequencing on Illumina platforms. These UDI are designed and validated to be used with <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">μChIPmentation for Histones</a> (Cat. No. C01011011), <a href="https://www.diagenode.com/en/p/chipmentation-kit-for-histones">ChIPmentation Kit for Histones</a> (Cat. No. C01011009), <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a> (Cat. No. C01011030), <a href="https://www.diagenode.com/en/categories/atac-seq">ATAC-seq Kit</a> (Cat. No. C01080002). The 24 UDI for tagmented libraries are compatible with other <b>tagmentation</b><b>-based library preparation </b>protocols, such as <a href="https://www.diagenode.com/en/categories/cutandtag">CUT&Tag</a> technologies.</p>
<p>3 sets of UDI for tagmented libraries are available:</p>
<p><strong>24 UDI for tagmented libraries - Set I</strong><br /> <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for tagmented libraries - Set II</a><br /><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3" target="_blank">24 UDI for tagmented libraries - Set III</a><br /><br /></p>
<p><span>Each set can be used for library multiplexing up to 24. All sets can be used simultaneously for library multiplexing up to 72.</span></p>
<p>Features:</p>
<ul>
<li>Multiplexing: <b>up to 72 samples </b>(using all 3 sets simultaneously)<b><br /></b></li>
<li>Allow for <b>identification of index hopping</b></li>
<li>Compatibility: <b>tagmentation</b><b>-based library preparation protocols</b></li>
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'info1' => '<p>The <b>24 UDI (Unique dual indexes) for </b><b>tagmented</b><b> libraries – Set I </b>is compatible with any <b>tagmentation</b><b>-based library preparation </b>protocols, such as <strong>ChIPmentation</strong>, <b>ATAC-seq</b> or <b>CUT&Tag</b> technologies.</p>
<p>The <b>24 UDI for </b><b>tagmented</b><b> libraries </b>provides combinations of barcodes where each barcode is uniquely attributed to one sample. This is a great tool to identify mistakes during index sequencing. A phenomenon, known as index hopping, can lead to misattribution of some reads to the wrong sample. This is particularly frequent with the NovaSeq6000, and thus the use of Unique Dual Indexing (UDI) is highly recommended when using this sequencer.</p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/UDI-for-tagmented-fig1.png" /></center>
<p></p>
<p><small><strong>Figure 1. Sequencing profiles of µChIPmentation libraries generated with 24 UDI for Tagmented libraries</strong> Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 UDI for Tagmented libraries – Set I (Cat. No. Cat. No. C01011034) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003) and rabbit IgG (Cat. No. C15410206) have been used. </small></p>
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'name' => 'Tagmentase (Tn5 transposase) - unloaded',
'description' => '<div class="extra-spaced"><center><img alt="Tagmentase (Tn5 transposase)" src="https://www.diagenode.com/img/banners/banner-tagmentase.jpg" caption="false" width="787" height="236" /></center></div>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
<p><a href="https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf" target="_blank">Protocol for transposome assembly</a></p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
</ul>
<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><img alt="Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1a.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="653" height="282" /></p>
<p><img alt="Tagmentase Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1b.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="645" height="278" /></p>
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<div class="small-12 medium-12 large-12 columns"><center><img alt="Tn5 transposase perfect for NGS" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure2.jpg" width="754" height="492" /></center></div>
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<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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'info2' => '<p><strong>Tagmentase (Tn5 transposase) - unloaded</strong></p>
<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
<p><strong>Storage conditions:</strong> Store at -20°C. Guaranteed stable for 6 months from date of receipt when stored properly.</p>
<p><strong>Storage buffer: </strong>Supplied in solution containing 50% v/v glycerol.</p>
<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones (developped for histone marks and some non-histone proteins), but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<br /> <a href="https://www.diagenode.com/files/application_notes/AN-iDealCUTandTag.pdf"><img src="https://www.diagenode.com/img/banners/cutandtag-appnote.png" /></a><br /><br /></div>
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<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
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<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
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<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
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<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
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<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
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<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
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<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p>目的のタンパク質のみをターゲットとすることが重要であるため、CUT＆Tagアッセイの成功には非常に特異的な抗体が不可欠です。 厳密な検証基準を満たしている特異性が検証されているDiagenode ChIP-seqグレードの抗体は、CUT＆Tagアッセイでの高いパフォーマンスを保証します。</p>
<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> <span>- compatible with histome marks and some transcription factors</span></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-a.jpg" width="600" height="265" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">B.<br /><center><img alt="pA-Tn5 Transposase unloaded" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-b.jpg" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> - compatible with histome marks and some transcription factors</li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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'description' => '<p><span>The negative Ctrl IgG from rabbit has been extensively validated in chromatin immunoprecipitation assays (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy animals. This IgG preparation is intended for use as a negative control in ChIP experiments for specific antibodies made in rabbit. The negative Ctrl IgG from rabbit should be used for ChIP in parallel with specific antibody at the same concentration as the specific antibody. </span></p>
<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-chip.jpg" alt="Rabbit IgG Antibody ChIP Grade" /></p>
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<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'info2' => '<p>The negative control IgG from rabbit has been extensively validated in chromatin immunoprecipitation (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy rabbits. This IgG preparation is intended for use as a negative control in ChIP, MeDIP, IF and other experiments performed with specific antibodies made in rabbit. The negative control IgG from rabbit should be used in parallel with the specific antibody at the same concentration. It is also included in many of our ChIP and MeDIP kits.</p>',
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'name' => '24 UDI for Tagmented libraries - Set I',
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<p>The <strong>24 UDI for tagmented libraries</strong> includes 24 primer pairs for unique dual-indexing allowing the multiplexing of up to <b>24 samples </b>for sequencing on Illumina platforms. These UDI are designed and validated to be used with <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">μChIPmentation for Histones</a> (Cat. No. C01011011), <a href="https://www.diagenode.com/en/p/chipmentation-kit-for-histones">ChIPmentation Kit for Histones</a> (Cat. No. C01011009), <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a> (Cat. No. C01011030), <a href="https://www.diagenode.com/en/categories/atac-seq">ATAC-seq Kit</a> (Cat. No. C01080002). The 24 UDI for tagmented libraries are compatible with other <b>tagmentation</b><b>-based library preparation </b>protocols, such as <a href="https://www.diagenode.com/en/categories/cutandtag">CUT&Tag</a> technologies.</p>
<p>3 sets of UDI for tagmented libraries are available:</p>
<p><strong>24 UDI for tagmented libraries - Set I</strong><br /> <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for tagmented libraries - Set II</a><br /><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3" target="_blank">24 UDI for tagmented libraries - Set III</a><br /><br /></p>
<p><span>Each set can be used for library multiplexing up to 24. All sets can be used simultaneously for library multiplexing up to 72.</span></p>
<p>Features:</p>
<ul>
<li>Multiplexing: <b>up to 72 samples </b>(using all 3 sets simultaneously)<b><br /></b></li>
<li>Allow for <b>identification of index hopping</b></li>
<li>Compatibility: <b>tagmentation</b><b>-based library preparation protocols</b></li>
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'info1' => '<p>The <b>24 UDI (Unique dual indexes) for </b><b>tagmented</b><b> libraries – Set I </b>is compatible with any <b>tagmentation</b><b>-based library preparation </b>protocols, such as <strong>ChIPmentation</strong>, <b>ATAC-seq</b> or <b>CUT&Tag</b> technologies.</p>
<p>The <b>24 UDI for </b><b>tagmented</b><b> libraries </b>provides combinations of barcodes where each barcode is uniquely attributed to one sample. This is a great tool to identify mistakes during index sequencing. A phenomenon, known as index hopping, can lead to misattribution of some reads to the wrong sample. This is particularly frequent with the NovaSeq6000, and thus the use of Unique Dual Indexing (UDI) is highly recommended when using this sequencer.</p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/UDI-for-tagmented-fig1.png" /></center>
<p></p>
<p><small><strong>Figure 1. Sequencing profiles of µChIPmentation libraries generated with 24 UDI for Tagmented libraries</strong> Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 UDI for Tagmented libraries – Set I (Cat. No. Cat. No. C01011034) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003) and rabbit IgG (Cat. No. C15410206) have been used. </small></p>
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'name' => 'Tagmentase (Tn5 transposase) - unloaded',
'description' => '<div class="extra-spaced"><center><img alt="Tagmentase (Tn5 transposase)" src="https://www.diagenode.com/img/banners/banner-tagmentase.jpg" caption="false" width="787" height="236" /></center></div>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
<p><a href="https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf" target="_blank">Protocol for transposome assembly</a></p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
</ul>
<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><img alt="Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1a.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="653" height="282" /></p>
<p><img alt="Tagmentase Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1b.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="645" height="278" /></p>
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<div class="small-12 medium-12 large-12 columns"><center><img alt="Tn5 transposase perfect for NGS" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure2.jpg" width="754" height="492" /></center></div>
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<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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'info2' => '<p><strong>Tagmentase (Tn5 transposase) - unloaded</strong></p>
<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
<p><strong>Storage conditions:</strong> Store at -20°C. Guaranteed stable for 6 months from date of receipt when stored properly.</p>
<p><strong>Storage buffer: </strong>Supplied in solution containing 50% v/v glycerol.</p>
<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones (developped for histone marks and some non-histone proteins), but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<br /> <a href="https://www.diagenode.com/files/application_notes/AN-iDealCUTandTag.pdf"><img src="https://www.diagenode.com/img/banners/cutandtag-appnote.png" /></a><br /><br /></div>
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<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
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<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
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<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
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<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
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<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
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<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
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<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p>目的のタンパク質のみをターゲットとすることが重要であるため、CUT＆Tagアッセイの成功には非常に特異的な抗体が不可欠です。 厳密な検証基準を満たしている特異性が検証されているDiagenode ChIP-seqグレードの抗体は、CUT＆Tagアッセイでの高いパフォーマンスを保証します。</p>
<p class="lead">CUT＆Tagアッセイに関する推奨製品はこちら</p>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064">loaded </a>or unloaded)</li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Negative CUT&Tag control <a href="https://www.diagenode.com/en/p/rabbit-igg-250-ug-250-ul">Rabbit IgG</a></li>
<li><i class="fa fa-arrow-circle-right"></i> Sequencing indexes <a href="https://www.diagenode.com/en/p/24-si-for-chipmentation">24 Single indexes</a></li>
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<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: <a href="https://www.diagenode.com/en/products/view/3064">loaded</a> or unloaded</span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag and similar assays (eg. TIP-seq)<br /></span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> <span>- compatible with histome marks and some transcription factors</span></li>
<li>pA/Tn5 Transposase (<a href="https://www.diagenode.com/en/products/view/3064" target="_blank">loaded </a>or unloaded)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-a.jpg" width="600" height="265" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">B.<br /><center><img alt="pA-Tn5 Transposase unloaded" src="https://www.diagenode.com/img/product/cutandtag/C01070002-fig-b.jpg" /></center></div>
</div>
<div class="row extra-spaced">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1: Quality control of pA-Tn5 transposase loaded with sequencing adapters</strong></p>
<p>A: The Fragment Analyzer trace showing the representative cleavage pattern of gDNA. The pA-Tn5 fusion protein (Cat. No. C01070002) loaded with sequencing adapters efficiently digests gDNA to a smear. 500 ng of human genomic DNA were incubated for 7 min at 55°C with 1 μl of pATn5 fusion protein loaded with appropriated adaptors in a tagmentation buffer (40mM Tris-HCl pH7.5, 40mM MgCl2 and 12.5% DMF). The reaction was stopped by adding SDS, cleaned-up and resolved on the Fragment Analyzer to assess the cleavage.</p>
<p>B: Representative screenshot at selected locus obtained using Diagenode pA-Tn5 fusion protein (Cat. No. C01070002) lloaded with sequencing adapters and H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) following CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)).</p>
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<p><strong>pA-Tn5 transposase</strong> is a fusion protein of hyperactive Tn5 transposase and protein A developed for the <strong>CUT&Tag assay</strong>. For convenience, the fusion protein is pre-loaded with sequencing adapters. Diagenode pA-Tn5 transposase is compatible with CUT&Tag and other antibody-tethered Tn5 based methods, like ACT-seq and CoBATCH.</p>
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<p><strong>pA-Tn5 Transposase</strong></p>
<ul style="list-style-type: circle;">
<li>High enrichment and specificity of the signal around the TSS</li>
<li>No contamination<span style="font-weight: 400;"> with </span><i><span style="font-weight: 400;">E. coli</span></i><span style="font-weight: 400;"> DNA</span></li>
<li><span style="font-weight: 400;">Flexible versions: loaded or <a href="https://www.diagenode.com/en/p/pa-tn5-transposase-unloaded">unloaded</a><br /></span></li>
<li><span style="font-weight: 400;">Excellent for CUT&Tag </span></li>
</ul>
<p>As it is crucial to target only the protein of interest, a highly specific antibody is critical for the success of the CUT&Tag assay. All Diagenode ChIP-seq grade antibodies are validated for specificity using strict validation criteria to assure <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">high performance in CUT&Tag</a>.</p>
<p class="lead"><strong>Check out all products recommended for CUT&Tag assay</strong>:</p>
<ul class="nobullet" style="list-style-type: circle;">
<li><a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24">iDeal CUT&Tag kit for Histones</a> - compatible with histome marks and some transcription factors</li>
<li>pA/Tn5 Transposase (loaded or <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">unloaded</a>)</li>
<li><a href="https://www.diagenode.com/en/applications/cut-and-tag">CUT&Tag grade</a> and <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade</a> antibodies</li>
<li>Negative & positive CUT&Tag controls: <span>Antibody Package for CUT&Tag: <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24">anti-rabbit</a> and <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24">anti-mouse</a></span></li>
<li>DNA purification: <a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2</a> or <a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></li>
<li>Sequencing indexes: <a href="https://www.diagenode.com/en/p/24-si-for-tagmented-libraries">single indexes</a> and <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">UDI</a></li>
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<div class="small-12 medium-12 large-12 columns">A.<br /><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-a.png" width="800" height="374" /></center><br /> B.<br /><center><img alt="pA-Tn5 Transposase loaded H3K9me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-b.png" width="800" height="374" /></center><br /> C.<br /><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation" src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-1-c.png" width="800" height="374" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 1.</strong> Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (A), <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">H3K9me3</a> (B) and <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (C) data obtained using Diagenode pA-Tn5 fusion protein (C01070001) and CUT&Tag protocol (Kaya-Okur, H.S., Nat Commun 10, 1930 (2019)) at selected loci. CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), H3K27me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>) and H3K9me3 polyclonal ChIP-seq grade antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k9me3-polyclonal-antibody-premium-50-mg">C15410193</a>).</p>
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<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K27me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-a.png" /></center></div>
<div class="small-12 medium-6 large-6 columns"><center><img alt="pA-Tn5 Transposase loaded H3K4me3 Validation " src="https://www.diagenode.com/img/product/cutandtag/pa-tn5-loaded-fig-2-b.png" /></center></div>
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<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2.</strong> CUT&Tag profiles show typical enrichments specific for a given histone mark. Representative screenshots for <a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">H3K27me3</a> (red) and <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">H3K4me3</a> (green) are shown at selected loci. H3K27me3, which marks inactive regions (red), does not show enrichment over open chromatin regions, marked by H3K4me3 (green). CUT&Tag was performed using 50,000 of K562 cells and Diagenode H3K4me3 polyclonal ChIP-seq antibody (Cat. No. <a href="https://www.diagenode.com/en/p/h3k4me3-polyclonal-antibody-premium-50-ug-50-ul">C15410003</a>), and H3K27me3 polyclonal ChIP-seq grade (Cat. No.<a href="https://www.diagenode.com/en/p/h3k27me3-polyclonal-antibody-premium-50-mg-27-ml">C15410195</a>).</p>
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'description' => '<p><span>The negative Ctrl IgG from rabbit has been extensively validated in chromatin immunoprecipitation assays (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy animals. This IgG preparation is intended for use as a negative control in ChIP experiments for specific antibodies made in rabbit. The negative Ctrl IgG from rabbit should be used for ChIP in parallel with specific antibody at the same concentration as the specific antibody. </span></p>
<p><span><span style="left: 94.4882px; top: 499.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.00223);">The negative Ctrl IgG is intended for use </span><span style="left: 94.4882px; top: 519.039px; font-size: 15px; font-family: sans-serif; transform: scaleX(1.02185);">as a negative control in ChIP, CUT&Tag, MeDIP, IF and other experiments performed with specific antibodies made in rabbit.</span></span></p>',
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-chip.jpg" alt="Rabbit IgG Antibody ChIP Grade" /></p>
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<p><small> <strong>Figure 1. ChIP with the Diagenode rabbit IgG negative control antibody</strong><br />ChIP assays were performed using the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) and the “iDeal ChIPseq” kit (Cat. No. C01010051) on sheared chromatin from 1 million HeLa cells. Rabbit IgG (cat. No. C15410206) was used as a negative IP control. One μg of antibody per ChIP experiment was used for both antibodies. Quantitative PCR was performed with primers specific for the promoters of the active GAPDH and EIF4A2 genes, and for the inactive MYOD1 gene and the Sat2 satellite repeat. Figure 1 shows the recovery, expressed as a % of input (the relative amount of immunoprecipitated DNA compared to input DNA after qPCR analysis).</small></p>
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<p><img src="https://www.diagenode.com/img/product/antibodies/C15410206-if.jpg" alt="Rabbit IgG validated in Immunofluorescence" /></p>
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<p><small> <strong>Figure 2. Immunofluorescence with the Diagenode rabbit IgG negative control antibody</strong><br />HeLa cells were stained with the Diagenode rabbit polyclonal antibody against H3K4me3 (Cat. No. C15410003) (top) and with DAPI. Rabbit IgG (Cat. No. C15410206) was used as a negative control (bottom). Cells were fixed with 4% formaldehyde for 10’ and blocked with PBS/TX-100 containing 5% normal goat serum and 1% BSA. The cells were immunofluorescently labeled with the H3K4me3 or rabbit IgG negative control antibody (left) diluted 1:200 in blocking solution followed by an anti-rabbit antibody conjugated to Alexa488. The middle panel shows staining of the nuclei with DAPI. A merge of the two stainings is shown on the right.</small></p>
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'info2' => '<p>The negative control IgG from rabbit has been extensively validated in chromatin immunoprecipitation (ChIP). It contains a spectrum of the IgG subclasses present in serum of healthy rabbits. This IgG preparation is intended for use as a negative control in ChIP, MeDIP, IF and other experiments performed with specific antibodies made in rabbit. The negative control IgG from rabbit should be used in parallel with the specific antibody at the same concentration. It is also included in many of our ChIP and MeDIP kits.</p>',
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'name' => '24 UDI for Tagmented libraries - Set I',
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<p>The <strong>24 UDI for tagmented libraries</strong> includes 24 primer pairs for unique dual-indexing allowing the multiplexing of up to <b>24 samples </b>for sequencing on Illumina platforms. These UDI are designed and validated to be used with <a href="https://www.diagenode.com/en/p/uchipmentation-for-histones-24-rxns">μChIPmentation for Histones</a> (Cat. No. C01011011), <a href="https://www.diagenode.com/en/p/chipmentation-kit-for-histones">ChIPmentation Kit for Histones</a> (Cat. No. C01011009), <a href="https://www.diagenode.com/en/p/tag-kit-for-chipmentation-24">TAG Kit for ChIPmentation</a> (Cat. No. C01011030), <a href="https://www.diagenode.com/en/categories/atac-seq">ATAC-seq Kit</a> (Cat. No. C01080002). The 24 UDI for tagmented libraries are compatible with other <b>tagmentation</b><b>-based library preparation </b>protocols, such as <a href="https://www.diagenode.com/en/categories/cutandtag">CUT&Tag</a> technologies.</p>
<p>3 sets of UDI for tagmented libraries are available:</p>
<p><strong>24 UDI for tagmented libraries - Set I</strong><br /> <a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set2">24 UDI for tagmented libraries - Set II</a><br /><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set3" target="_blank">24 UDI for tagmented libraries - Set III</a><br /><br /></p>
<p><span>Each set can be used for library multiplexing up to 24. All sets can be used simultaneously for library multiplexing up to 72.</span></p>
<p>Features:</p>
<ul>
<li>Multiplexing: <b>up to 72 samples </b>(using all 3 sets simultaneously)<b><br /></b></li>
<li>Allow for <b>identification of index hopping</b></li>
<li>Compatibility: <b>tagmentation</b><b>-based library preparation protocols</b></li>
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'info1' => '<p>The <b>24 UDI (Unique dual indexes) for </b><b>tagmented</b><b> libraries – Set I </b>is compatible with any <b>tagmentation</b><b>-based library preparation </b>protocols, such as <strong>ChIPmentation</strong>, <b>ATAC-seq</b> or <b>CUT&Tag</b> technologies.</p>
<p>The <b>24 UDI for </b><b>tagmented</b><b> libraries </b>provides combinations of barcodes where each barcode is uniquely attributed to one sample. This is a great tool to identify mistakes during index sequencing. A phenomenon, known as index hopping, can lead to misattribution of some reads to the wrong sample. This is particularly frequent with the NovaSeq6000, and thus the use of Unique Dual Indexing (UDI) is highly recommended when using this sequencer.</p>
<p></p>
<center><img src="https://www.diagenode.com/img/product/kits/UDI-for-tagmented-fig1.png" /></center>
<p></p>
<p><small><strong>Figure 1. Sequencing profiles of µChIPmentation libraries generated with 24 UDI for Tagmented libraries</strong> Chromatin preparation and immunoprecipitation have been performed on 10.000 cells using the µChIPmentation Kit for Histones (Cat. No. C01011011) and 24 UDI for Tagmented libraries – Set I (Cat. No. Cat. No. C01011034) using K562 cells. The Diagenode antibodies targeting H3K4me3 (Cat. No. C15410003) and rabbit IgG (Cat. No. C15410206) have been used. </small></p>
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'name' => 'Tagmentase (Tn5 transposase) - unloaded',
'description' => '<div class="extra-spaced"><center><img alt="Tagmentase (Tn5 transposase)" src="https://www.diagenode.com/img/banners/banner-tagmentase.jpg" caption="false" width="787" height="236" /></center></div>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
<p><a href="https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf" target="_blank">Protocol for transposome assembly</a></p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
</ul>
<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><img alt="Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1a.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="653" height="282" /></p>
<p><img alt="Tagmentase Tn5 transposase" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure-1b.jpg" style="display: block; margin-left: auto; margin-right: auto;" width="645" height="278" /></p>
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<div class="small-12 medium-12 large-12 columns"><center><img alt="Tn5 transposase perfect for NGS" src="https://www.diagenode.com/img/product/reagents/tagmentase-figure2.jpg" width="754" height="492" /></center></div>
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<div class="row">
<div class="small-12 medium-12 large-12 columns">
<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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'info2' => '<p><strong>Tagmentase (Tn5 transposase) - unloaded</strong></p>
<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
<p><strong>Storage conditions:</strong> Store at -20°C. Guaranteed stable for 6 months from date of receipt when stored properly.</p>
<p><strong>Storage buffer: </strong>Supplied in solution containing 50% v/v glycerol.</p>
<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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<p><strong>CUT&Tag</strong>-sequencing (<strong>C</strong>leavage <strong>U</strong>nder <strong>T</strong>argets and <strong>Tag</strong>mentation) is a new alternative method to ChIP-seq combining antibody-targeted controlled cleavage by a protein A-Tn5 fusion with massively parallel DNA sequencing to identify the binding sites of DNA-associated proteins. At Diagenode we offer a complete solution for CUT&Tag – our iDeal CUT&Tag for Histones (developped for histone marks and some non-histone proteins), but also stand-alone fusion protein – pA-Tn5 Transposase. Moreover, we have validated our <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies">ChIP-seq grade antibodies</a> in <a href="https://www.diagenode.com/en/categories/cut-and-tag-antibodies">CUT&Tag</a> proving their high performance in this assay.</p>
<br /> <a href="https://www.diagenode.com/files/application_notes/AN-iDealCUTandTag.pdf"><img src="https://www.diagenode.com/img/banners/cutandtag-appnote.png" /></a><br /><br /></div>
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<li class="accordion-navigation"><a href="#v5" style="color: #13b29c;"><i class="fa fa-caret-right"></i> How does it work?</a>
<div id="v5" class="content">
<p>The iDeal CUT&Tag protocol involves the binding of cells on a solid phase ConA magnetic beads, allowing magnetic handling of the cells for the major steps of the protocol. Bead-bound cells are permeabilized, incubated with primary antibody against a target of interest and secondary antibody. Then, Diagenode’s protein pA-Tn5 Transposase - loaded is bound to the complex. Protein A guides Tn5 transposase on chromatin to the antibody attached to its target. Tn5 transposase is activated by Mg+2 ions to insert the sequencing adaptors into genomic regions of interest. DNA is then purified and the tagmented genomic regions of interest are amplified by PCR using Diagenode’s Primer Indexes for tagmented libraries.</p>
<img src="https://www.diagenode.com/img/product/kits/workflow-cutandtag.jpg" /></div>
<h2>Products for CUT&Tag assay</h2>
<h3 class="diacol">Complete solutions</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/ideal-cut-and-tag-kit-for-histones-24" target="_blank">iDeal CUT&Tag kit for Histones</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
</ul>
<h3 class="diacol">Fusion protein</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3064" target="_blank">pA/Tn5 Transposase (loaded)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/products/view/3065" target="_blank">pA/Tn5 Transposase (unloaded)</a></li>
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<h3 class="diacol">CUT&Tag grade antibodies</h3>
<ul class="nobullet">
<li>Antibodies <a href="https://www.diagenode.com/en/applications/cut-and-tag">validated in CUT&Tag</a></li>
<li>Check out our list of <a href="https://www.diagenode.com/en/categories/chip-seq-grade-antibodies" target="_blank">ChIP-seq grade antibodies</a></li>
<li>Read more about the performance of Diagenode antibodies in <a href="https://www.diagenode.com/en/pages/cut-and-tag" target="_blank">CUT&Tag</a></li>
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<h3 class="diacol">Positive & Negative CUT&Tag control</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antirabbit-24" target="_blank">Antibody package for CUT&Tag (anti-rabbit)</a></li>
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/p/antibody-package-cut-and-tag-antimouse-24" target="_blank">Antibody package for CUT&Tag (anti-mouse)</a></li>
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<h3 class="diacol">DNA purification</h3>
<p style="padding-left: 30px;"><a href="https://www.diagenode.com/en/p/ipure-kit-v2-x24">IPure kit v2<br /></a><a href="https://www.diagenode.com/en/p/microchip-diapure-columns-50-rxns">MicroChIP DiaPure columns</a></p>
<h3 class="diacol">Sequencing indexes</h3>
<ul class="nobullet">
<li><i class="fa fa-arrow-circle-right"></i> <a href="https://www.diagenode.com/en/categories/primer-indexes-for-tagmented-libraries" target="_blank">Primer indexes for tagmented libraries</a></li>
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<p>Diagenode Tagmentase is a hyperactive Tn5 transposase with the potential to enhance epigenetic studies. Its ability to cut DNA and insert sequences of interest in one step makes it the perfect companion for Next-Generation Sequencing experiments using powerful technologies such as ATAC-seq, ChIPmentation, CHANGE-seq and other. The enzyme is not loaded with DNA oligos, providing flexibility of application. To ensure optimal results the concentration may be adjusted with Diagenode <a href="https://www.diagenode.com/en/p/tagmentase-dilution-buffer">Tagmentase Dilution Buffer</a> (Cat. No. C01070011), available separately.</p>
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<p>Using Diagenode’s Tagmentase (Tn5 transposase) you may also need:</p>
<ul>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-1x-1ml">Tagmentation Buffer (1x)</a></li>
<li><a href="https://www.diagenode.com/en/p/tagmentation-buffer-2x">Tagmentation Buffer (2x)</a></li>
<li><a href="https://www.diagenode.com/en/p/24-unique-dual-indexes-for-tagmented-libraries-set1">24 UDI for tagmented libraries</a></li>
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<p>Looking for loaded Tagmentase? Please go to <a href="https://www.diagenode.com/en/p/tagmentase-loaded-30">Tagmentase (Tn5 transposase) - loaded</a>.</p>',
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<p><strong>Figure 1: Efficient fragmentation of the lambda DNA after incubation with the Tagmentase</strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with diluted Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). Profiles show the size of lambda DNA before (A) and after treatment with Tagmentase (B).</p>
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<p><strong>Figure 2: Fragmentation efficiency depending on the amount of Tagmentase </strong><br />For fragmentation, 100 ng of DNA from bacteriophage lambda were incubated with Diagenode Tagmentase (Cat. No. C01070010) and Tagmentation buffer (1x) (Cat. No. C01019042) for 7 min at 55°C. The Tagmentase was previously diluted with the Tagmentase Dilution Buffer (Cat. No.) at ¼ and 1/16 dilutions. The reaction was stopped by addition of SDS (0.2% final concentration). After clean-up using AMPure XP beads (Beckman Coulter) on Diagenode IP-Star robot, the size of the DNA was assessed on Fragment Analyzer (Agilent), using the HS Large Fragment 50kb Kit (Agilent). The migration of the samples shows variations of the size distribution according to the amount of Tagmentase used for the reaction.</p>
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<div><span style="font-family: inherit;">Protein Molecular weight: 53.3 kDa</span></div>
<p>Expressed: in Escherichia coli</p>
<p><strong>Product description:</strong> Diagenode Tagmentase – unloaded is a hyperactive Tn5 transposase. The enzyme catalyzes “cut and paste” tagmentation reaction and can be used to insert any target DNA in vitro.</p>
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<p><strong>Properties & Usage: </strong>The enzyme should be loaded with appropriate oligonucleotides prior to use. An efficient transposition require that insert DNA have a specific 19-bp transposase recognition sequence (Mosaic End or ME sequence) at each of its ends. The transposome assembly protocol can be found at https://www.diagenode.com/files/protocols/PRO-Transposome-Assembly-V2.pdf Tagmentase is dependent on Mg++ for activity. Avoid chelators, such as EDTA/EGTA, in reaction buffers. The enzyme is active at pH 7.5-8 at 37-55°C. SDS, EDTA/EGTA or heating to 65°C will inactivate the enzyme.</p>
<p><strong>Applications: </strong>Tagmentase (Tn5 transposase) – unloaded can be used in a variety of applications including transgenic experiments, barcoding and library construction for second-generation sequencing. Please note that an additional optimization might be required for custom protocols including the enzyme dose- and time-response experiments.</p>
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