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<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<p><strong>Figure 1. Optimal chromatin shearing profile</strong><br /> HeLa cells were fixed with formaldehyde for 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.</p>
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<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<p><strong>Figure 1. Optimal chromatin shearing profile</strong><br /> HeLa cells were fixed with formaldehyde for 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.</p>
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<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<p><strong>Cell lines</strong>:</p>
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<li>Human: A549, A673, BT-549, CD4 T, HCC1806, HeLa, HepG2, HFF, HK-GFP-MR, ILC, K562, KYSE-180, LapC4, M14, MCF7, MDA-MB-231, MDA-MB-436, RDES, SKNO1, VCaP, U2-OS, ZR-75-1</li>
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<li>Cattle: pbMEC, <span>MAC-T</span></li>
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<li>Mouse: kidney, heart, brain, iris, liver, limbs from E10.5 embryos</li>
<li>Horse: liver, brain, heart, lung, skeletal muscle, lamina, ovary</li>
<li>Other tissues: compatible, not tested</li>
</ul>
<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
<div class="small-12 medium-12 large-12 columns text-center"><br /><img src="https://www.diagenode.com/img/applications/pico_dna_shearing_fig2.png" /></div>
<div class="small-10 medium-10 large-10 columns end small-offset-1"><small> <br /><strong>Panel A, 10 µl volume:</strong> Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). <strong>Panel B, 100 µl volume:</strong> Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). <strong>Panel C, 300 µl volume:</strong> Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).</small></div>
<div class="small-12 medium-12 large-12 columns"><br /><br /></div>
<div class="small-12 medium-12 large-12 columns">
<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<div class="small-12 medium-12 large-12 columns">
<div class="page" title="Page 7">
<table>
<tbody>
<tr valign="middle">
<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
</table>
<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
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'description' => '<p>Optimal detergent concentration is required for best results with chromatin shearing. Diagenode offers four kits with different SDS concentrations that have been pre-optimized for your specific workflow requirements. Our Chromatin EasyShear Kits (previous name: Chromatin Shearing Optimization Kits) together with the Bioruptor combine efficient cell lysis and chromatin shearing leading to consistent results.</p>
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<li>The optimization of the chromatin shearing and/or chromatin preparation prior to any ChIP protocol</li>
<li>The optimization of the chromatin shearing of a new cell line/new sample type prior to ChIP using Diagenode’s ChIP kits</li>
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<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
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<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<p><strong>Figure 1. Optimal chromatin shearing profile</strong><br /> HeLa cells were fixed with formaldehyde for 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.</p>
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<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/product/kits/chromatin-easyshear-low-sds-fig-2a.jpg" /> <img src="https://www.diagenode.com/img/product/kits/chromatin-easyshear-low-sds-fig-2b.jpg" /></div>
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<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<li>Other tissues: compatible, not tested</li>
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<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
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<div class="small-10 medium-10 large-10 columns end small-offset-1"><small> <br /><strong>Panel A, 10 µl volume:</strong> Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). <strong>Panel B, 100 µl volume:</strong> Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). <strong>Panel C, 300 µl volume:</strong> Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).</small></div>
<div class="small-12 medium-12 large-12 columns"><br /><br /></div>
<div class="small-12 medium-12 large-12 columns">
<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<div class="page" title="Page 7">
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<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
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<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
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<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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'description' => '<p>Old name of the kit: <strong>Chromatin shearing optimization kit</strong> – <strong>Low SDS (iDeal Kit for TFs)</strong></p>
<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<ul>
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<li>SDS concentration optimized for the workflow for TFs</li>
<li>Validated for cells, tissue and FFPE samples</li>
<li>Preserves the epitopes</li>
<li>Validated with the Bioruptor ultrasonicator</li>
<li>Quality of chromatin sample confirmed by ChIP-seq</li>
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<div class="small-9 medium-9 large-9 columns">
<p><strong>Figure 1. Optimal chromatin shearing profile</strong><br /> HeLa cells were fixed with formaldehyde for 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.</p>
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<div class="small-4 medium-4 large-4 columns"><img src="https://www.diagenode.com/img/product/kits/chromatin-easyshear-low-sds-fig-2a.jpg" /> <img src="https://www.diagenode.com/img/product/kits/chromatin-easyshear-low-sds-fig-2b.jpg" /></div>
<div class="small-8 medium-8 large-8 columns">
<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<p><strong>Cell lines</strong>:</p>
<ul>
<li>Human: A549, A673, BT-549, CD4 T, HCC1806, HeLa, HepG2, HFF, HK-GFP-MR, ILC, K562, KYSE-180, LapC4, M14, MCF7, MDA-MB-231, MDA-MB-436, RDES, SKNO1, VCaP, U2-OS, ZR-75-1</li>
<li>Mouse: ESC, NPCs, BZ, GT1-7, acinar cells, HSPCs, Th2 cells, keratinocytes</li>
<li>Cattle: pbMEC, <span>MAC-T</span></li>
<li>Other cell lines / species: compatible, not tested</li>
</ul>
<p><strong>Tissues</strong>:</p>
<ul>
<li>Mouse: kidney, heart, brain, iris, liver, limbs from E10.5 embryos</li>
<li>Horse: liver, brain, heart, lung, skeletal muscle, lamina, ovary</li>
<li>Other tissues: compatible, not tested</li>
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<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<p><strong>Figure 1. Optimal chromatin shearing profile</strong><br /> HeLa cells were fixed with formaldehyde for 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.</p>
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<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<p><strong>Cell lines</strong>:</p>
<ul>
<li>Human: A549, A673, BT-549, CD4 T, HCC1806, HeLa, HepG2, HFF, HK-GFP-MR, ILC, K562, KYSE-180, LapC4, M14, MCF7, MDA-MB-231, MDA-MB-436, RDES, SKNO1, VCaP, U2-OS, ZR-75-1</li>
<li>Mouse: ESC, NPCs, BZ, GT1-7, acinar cells, HSPCs, Th2 cells, keratinocytes</li>
<li>Cattle: pbMEC, <span>MAC-T</span></li>
<li>Other cell lines / species: compatible, not tested</li>
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<p><strong>Tissues</strong>:</p>
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<li>Mouse: kidney, heart, brain, iris, liver, limbs from E10.5 embryos</li>
<li>Horse: liver, brain, heart, lung, skeletal muscle, lamina, ovary</li>
<li>Other tissues: compatible, not tested</li>
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<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
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<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
</tr>
</tbody>
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<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
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'description' => '<p>Optimal detergent concentration is required for best results with chromatin shearing. Diagenode offers four kits with different SDS concentrations that have been pre-optimized for your specific workflow requirements. Our Chromatin EasyShear Kits (previous name: Chromatin Shearing Optimization Kits) together with the Bioruptor combine efficient cell lysis and chromatin shearing leading to consistent results.</p>
<p>The Chromatin EasyShear Kits are recommended for:</p>
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<li>The optimization of the chromatin shearing of a new cell line/new sample type prior to ChIP using Diagenode’s ChIP kits</li>
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<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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'description' => '<p>X-linked myotubular myopathy (XLMTM) is a fatal neuromuscular disorder caused by loss of function mutations in MTM1. At present, there are no directed therapies for XLMTM, and incomplete understanding of disease pathomechanisms. To address these knowledge gaps, we performed a drug screen in mtm1 mutant zebrafish and identified four positive hits, including valproic acid, which functions as a potent suppressor of the mtm1 zebrafish phenotype via HDAC inhibition. We translated these findings to a mouse XLMTM model, and showed that valproic acid ameliorates the murine phenotype. These observations led us to interrogate the epigenome in Mtm1 knockout mice; we found increased DNA methylation, which is normalized with valproic acid, and likely mediated through aberrant 1-carbon metabolism. Finally, we made the unexpected observation that XLMTM patients share a distinct DNA methylation signature, suggesting that epigenetic alteration is a conserved disease feature amenable to therapeutic intervention.</p>',
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'description' => '<p>Head and Neck Squamous Cell Carcinoma (HNSCC) is a heterogeneous disease with relatively high morbidity and mortality rates. The lack of effective therapies, high recurrence rates and drug resistance driven in part, by tumor heterogeneity, contribute to the poor prognosis for patients diagnosed with this cancer. This problem is further exacerbated by the fact that key regulatory factors contributing to the disease diversity remains largely elusive. Here, we have identified EHF as an important member of the ETS family of transcription factors that is highly expressed in normal oral tissues, but lost during HNSCC progression. Interestingly, HNSCC tumors and cell lines exhibited a dichotomy of high and low EHF expression, and patients whose tumors retained EHF expression showed significantly better prognosis, suggesting a potential tumor suppressive role for EHF. To address this, we have performed gain and loss of function studies and leveraged bulk and single-cell cancer genomic datasets to identify global EHF targets by RNA-sequencing (RNA-seq) and Chromatin Immunoprecipitation and next generation sequencing (ChIP-seq) experiments of HNSCC cell lines. These mechanistic studies have revealed that EHF, acts as a regulator of a broad spectrum of metabolic processes, specifically targeting regulators of redox homeostasis such as NRF2 and SOX2. Our immunostaining results confirm the mutually exclusive expression patterns of EHF and SOX2 in HNSCC tumors and suggest a possible role for these two factors in establishing discrete metabolic states within the tumor microenvironment. Taken together, EHF may serve as a novel prognostic marker for classifying HNSCC patients for actionable and targeted therapeutic intervention.</p>',
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
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<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<p><strong>Figure 1. Optimal chromatin shearing profile</strong><br /> HeLa cells were fixed with formaldehyde for 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.</p>
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<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<p>A high quality chromatin preparation is very complex and requires a lot of optimization. Chromatin EasyShear Kit – Low SDS is an optimized solution for efficient chromatin preparation prior to ChIP. The protocol, buffers composition, SDS concentration (0.2%) is optimized for the preparation of chromatin prior to ChIP on <strong>transcription factors and other non-histone proteins</strong>. The kit has been validated with the Bioruptor ultrasonicator for efficient chromatin shearing, leading to chromatin fragments <strong>suitable for ChIP</strong> with the preserved <strong>epitopes</strong>. The kit is validated for cells, tissues and FFPE samples.</p>
<p>Check out all of the Chromatin EasyShear Kits.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a>Read more</a></p>
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<p><strong>Figure 1. Optimal chromatin shearing profile</strong><br /> HeLa cells were fixed with formaldehyde for 10 min and chromatin was prepared according to Diagenode’s Chromatin EasyShear Kit - Low SDS (Cat. No. C01020013). Samples were sonicated for 5-10-15 cycles of 30” ON/30” OFF as indicated with Bioruptor Pico using 1.5 ml Bioruptor microtubes with caps (Cat. No. C30010016) followed by de-crosslinking and DNA purification. The fragment size was assessed using agarose gel electrophoresis. A 100 bp ladder was loaded as the size standard.</p>
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<p><strong>Figure 2. Chromatin precipitation</strong><br /> Sheared chromatin (obtained with the Chromatin EasyShear Kit – Low SDS) has been used for immunoprecipitation with CTCF and IgG (negative control) antibodies. Quantitative PCR was performed with positive (H19) and negative (Myoglobine exon 2) control regions. The Figure 2 shows the recovery expressed as % of input (the relative amount of immunoprecipitated DNA compared to input DNA (panel A) and as enrichment fold of positive locus over negative (panel B).</p>
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<p><strong>Cell lines</strong>:</p>
<ul>
<li>Human: A549, A673, BT-549, CD4 T, HCC1806, HeLa, HepG2, HFF, HK-GFP-MR, ILC, K562, KYSE-180, LapC4, M14, MCF7, MDA-MB-231, MDA-MB-436, RDES, SKNO1, VCaP, U2-OS, ZR-75-1</li>
<li>Mouse: ESC, NPCs, BZ, GT1-7, acinar cells, HSPCs, Th2 cells, keratinocytes</li>
<li>Cattle: pbMEC, <span>MAC-T</span></li>
<li>Other cell lines / species: compatible, not tested</li>
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<p><strong>Tissues</strong>:</p>
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<li>Mouse: kidney, heart, brain, iris, liver, limbs from E10.5 embryos</li>
<li>Horse: liver, brain, heart, lung, skeletal muscle, lamina, ovary</li>
<li>Other tissues: compatible, not tested</li>
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<p>Did you use the iDeal ChIP-seq for Transcription Factors Kit on other cell line / tissue / species? <a href="mailto:agnieszka.zelisko@diagenode.com?subject=Species, cell lines, tissues tested with the iDeal ChIP-seq Kit for TF&body=Dear Customer,%0D%0A%0D%0APlease, leave below your feedback about the iDeal ChIP-seq for Transcription Factors (cell / tissue type, species, other information...).%0D%0A%0D%0AThank you for sharing with us your experience !%0D%0A%0D%0ABest regards,%0D%0A%0D%0AAgnieszka Zelisko-Schmidt, PhD">Let us know!</a></p>
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<div class="small-12 medium-12 large-12 columns">The most important steps for a successful ChIP include both cell fixation and lysis, and chromatin shearing. Researchers often overlook the critical nature of both of these steps. Eliminating inconsistencies in the shearing step, <strong>Diagenode's Bioruptor</strong><sup>®</sup> uses state-of-the-art ultrasound <strong>ACT</strong> (<strong>A</strong>daptive <strong>C</strong>avitation <strong>T</strong>echnology) to efficiently shear chromatin. ACT enables the highest chromatin quality for high IP efficiency and sensitivity for ChIP experiments with gentle yet highly effective shearing forces. Additionally, the Bioruptor<sup>®</sup> provides a precisely controlled temperature environment that preserves chromatin from heat degradation such that protein-DNA complexes are well-preserved for sensitive, unbiased, and accurate ChIP.<br /><br /> <strong>Diagenode's Bioruptor</strong><sup>®</sup> is the instrument of choice for chromatin shearing used for a number of downstream applications such as qPCR and ChIP-seq that require optimally sheared, unbiased chromatin.</div>
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<div class="small-10 medium-10 large-10 columns end small-offset-1"><small> <br /><strong>Panel A, 10 µl volume:</strong> Chromatin samples are sheared for 10, 20 and 30 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.1 ml Bioruptor® Microtubes (Cat. No. B01200041). <strong>Panel B, 100 µl volume:</strong> Chromatin samples are sheared for 10 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using 0.65 ml Bioruptor® Microtubes (Cat. No. WA-005-0500). <strong>Panel C, 300 µl volume:</strong> Chromatin samples are sheared for 5, 10 and 15 cycles of 30 sec ON/30 sec OFF with the Bioruptor® Pico using using 1.5 ml Bioruptor microtubes (Cat. No. C30010016). Prior to de-crosslinking, samples are treated with RNase cocktail mixture at 37°C during 1 hour. The sheared chromatin is then de-crosslinked overnight and phenol/chloroform purified as described in the kit manual. 10 µl of DNA (equivalent of 500, 000 cells) are analyzed on a 2% agarose gel (MW corresponds to the 100 bp DNA molecular weight marker).</small></div>
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<div class="small-12 medium-12 large-12 columns">
<p>It is important to establish optimal conditions to shear crosslinked chromatin to get the correct fragment sizes needed for ChIP. Usually this process requires both optimizing sonication conditions as well as optimizing SDS concentration, which is laborious. With the Chromatin Shearing Optimization Kits, optimization is fast and easy - we provide optimization reagents with varying concentrations of SDS. Moreover, our Chromatin Shearing Optimization Kits can be used for the optimization of chromatin preparation with our kits for ChIP.</p>
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<td></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-100-million-cells">Chromatin Shearing Kit Low SDS (for Histone)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-low-sds-for-tfs-25-rxns">Chromatin Shearing Kit Low SDS (for TF)</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-high-sds-100-million-cells">Chromatin Shearing Kit High SDS</a></strong></td>
<td style="text-align: center;"><strong><a href="https://www.diagenode.com/p/chromatin-shearing-optimization-kit-medium-sds-100-million-cells">Chromatin Shearing Kit (for Plant)</a></strong></td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>SDS concentration</strong></p>
</td>
<td style="text-align: center;">
<p>< 0.1%</p>
</td>
<td style="text-align: center;">
<p>0.2%</p>
</td>
<td style="text-align: center;">
<p>1%</p>
</td>
<td style="text-align: center;">
<p>0.5%</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Nuclei isolation</strong></p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
<td style="text-align: center;">
<p>No</p>
</td>
<td style="text-align: center;">
<p>Yes</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Allows for shearing of... cells/tissue</strong></p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>100 million cells</p>
</td>
<td style="text-align: center;">
<p>up to 25 g of tissue</p>
</td>
</tr>
<tr valign="middle" style="background-color: #fff;">
<td>
<p><strong>Corresponding to shearing buffers from</strong></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-x24-24-rxns">iDeal ChIP-seq kit for Histones</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/ideal-chip-seq-kit-for-transcription-factors-x24-24-rxns">iDeal ChIP-seq Kit for Transcription Factors</a></p>
<p><a href="https://www.diagenode.com/en/p/ideal-chip-qpcr-kit">iDeal ChIP qPCR kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/true-microchip-kit-x16-16-rxns">True MicroChIP kit</a></p>
</td>
<td style="text-align: center;">
<p><a href="https://www.diagenode.com/en/p/universal-plant-chip-seq-kit-x24-24-rxns">Universal Plant ChIP-seq kit</a></p>
</td>
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<p><em><span style="font-weight: 400;">Table comes from our </span><a href="https://www.diagenode.com/protocols/bioruptor-pico-chromatin-preparation-guide"><span style="font-weight: 400;">Guide for successful chromatin preparation using the Bioruptor® Pico</span></a></em></p>
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'description' => '<p>Optimal detergent concentration is required for best results with chromatin shearing. Diagenode offers four kits with different SDS concentrations that have been pre-optimized for your specific workflow requirements. Our Chromatin EasyShear Kits (previous name: Chromatin Shearing Optimization Kits) together with the Bioruptor combine efficient cell lysis and chromatin shearing leading to consistent results.</p>
<p>The Chromatin EasyShear Kits are recommended for:</p>
<ul>
<li>The optimization of the chromatin shearing and/or chromatin preparation prior to any ChIP protocol</li>
<li>The optimization of the chromatin shearing of a new cell line/new sample type prior to ChIP using Diagenode’s ChIP kits</li>
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<p>Check all <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</p>
<p>Guide for the optimal chromatin preparation using Chromatin EasyShear Kits – <a href="https://www.diagenode.com/en/pages/chromatin-prep-easyshear-kit-guide">Read more</a></p>
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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'description' => '<p>Head and Neck Squamous Cell Carcinoma (HNSCC) is a heterogeneous disease with relatively high morbidity and mortality rates. The lack of effective therapies, high recurrence rates and drug resistance driven in part, by tumor heterogeneity, contribute to the poor prognosis for patients diagnosed with this cancer. This problem is further exacerbated by the fact that key regulatory factors contributing to the disease diversity remains largely elusive. Here, we have identified EHF as an important member of the ETS family of transcription factors that is highly expressed in normal oral tissues, but lost during HNSCC progression. Interestingly, HNSCC tumors and cell lines exhibited a dichotomy of high and low EHF expression, and patients whose tumors retained EHF expression showed significantly better prognosis, suggesting a potential tumor suppressive role for EHF. To address this, we have performed gain and loss of function studies and leveraged bulk and single-cell cancer genomic datasets to identify global EHF targets by RNA-sequencing (RNA-seq) and Chromatin Immunoprecipitation and next generation sequencing (ChIP-seq) experiments of HNSCC cell lines. These mechanistic studies have revealed that EHF, acts as a regulator of a broad spectrum of metabolic processes, specifically targeting regulators of redox homeostasis such as NRF2 and SOX2. Our immunostaining results confirm the mutually exclusive expression patterns of EHF and SOX2 in HNSCC tumors and suggest a possible role for these two factors in establishing discrete metabolic states within the tumor microenvironment. Taken together, EHF may serve as a novel prognostic marker for classifying HNSCC patients for actionable and targeted therapeutic intervention.</p>',
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<p><span>The first critical step of a successful Chromatin immunoprecipitation (ChIP) experiment is the preparation of sheared chromatin. We therefore suggest to use one of our optimized shearing ChIP kits.</span></p>
<span>• Chromatin EasyShear Kit - Ultra Low SDS <br />• Chromatin EasyShear Kit - Low SDS <br />• Chromatin EasyShear Kit - High SDS </span></div>
<div class="column"><span>• Chromatin EasyShear Kit for Plant</span><span></span>
<p><span></span></p>
<p><span>Our Chromatin EasyShear kits are used in combination with the Bioruptor</span><span>® </span><span>in order to ensure a highly reproducible chromatin shearing and make sure you obtain the right fragment size needed for your experiment. Establish optimal conditions required for shearing cross-linked chromatin (protein-DNA) is usually laborious; the protocol of the Chromatin EasyShear kits is fast, easy-to-use and optimized to get the best results possible. </span></p>
<p><span>Learn more about Diagenode <a href="https://www.diagenode.com/en/categories/chromatin-shearing">Chromatin EasyShear Kits</a>.</span></p>
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include - APP/View/Products/view.ctp, line 755
View::_evaluate() - CORE/Cake/View/View.php, line 971
View::_render() - CORE/Cake/View/View.php, line 933
View::render() - CORE/Cake/View/View.php, line 473
Controller::render() - CORE/Cake/Controller/Controller.php, line 963
ProductsController::slug() - APP/Controller/ProductsController.php, line 1052
ReflectionMethod::invokeArgs() - [internal], line ??
Controller::invokeAction() - CORE/Cake/Controller/Controller.php, line 491
Dispatcher::_invoke() - CORE/Cake/Routing/Dispatcher.php, line 193
Dispatcher::dispatch() - CORE/Cake/Routing/Dispatcher.php, line 167
[main] - APP/webroot/index.php, line 118
×