| Datasheet MethylTaq Plus 2X Master Mix TECHNICAL DATASHEET Datasheet MethylTaq Plus 2X Master Mix | Download |
MethylTaq Plus 2X Master Mix is a ready-to-use cocktail containing all components required for the amplification of bisulfite converted NGS libraries, except primers and template. This convenient 2X Master Mix contains MethylTaq Plus DNA polymerase, each dNTP (dATP, dCTP, dGTP, dTTP, but not dUTP) [0.3 mM], MgCl2 [2.5 mM] and reaction buffer.
| Datasheet MethylTaq Plus 2X Master Mix TECHNICAL DATASHEET Datasheet MethylTaq Plus 2X Master Mix | Download |
| MethylTaq Plus 2X Master Mix SDS GB en | Download |
| MethylTaq Plus 2X Master Mix SDS US en | Download |
| MethylTaq Plus 2X Master Mix SDS DE de | Download |
| MethylTaq Plus 2X Master Mix SDS JP ja | Download |
| MethylTaq Plus 2X Master Mix SDS BE nl | Download |
| MethylTaq Plus 2X Master Mix SDS FR fr | Download |
| MethylTaq Plus 2X Master Mix SDS BE fr | Download |
| MethylTaq Plus 2X Master Mix SDS ES es | Download |
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Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p> </div> <div class="small-12 medium-4 large-4 columns text-center"><a href="../landing-pages/dna-methylation-grant-applications"><img src="https://www.diagenode.com/img/banners/banner-dna-grant.png" alt="" /></a></div> <div style="text-align: justify;" class="small-12 medium-12 large-12 columns"> <p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p> </div> <div class="small-12 medium-12 large-12 columns"><br /><br /> <ul class="accordion" data-accordion=""> <li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a> <div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base. <p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p> </div> </li> </ul> <br /> <h2>Main DNA methylation technologies</h2> <p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p> <div class="row"> <ol> <li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li> </ol> <p><span style="font-weight: 400;"> </span></p> <div class="row"> <table> <thead> <tr> <th></th> <th>Description</th> <th width="350">Features</th> </tr> </thead> <tbody> <tr> <td><strong>Bisulfite conversion</strong></td> <td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><b>Methylated DNA enrichment</b></td> <td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><strong>Restriction enzyme-based digestion</strong></td> <td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li> </ul> </td> </tr> </tbody> </table> </div> </div> <div class="row"></div> </div> </div> <div class="large-12 columns"></div> </div>', 'in_footer' => true, 'in_menu' => true, 'online' => true, 'tabular' => true, 'slug' => 'epigenetics-dna-methylation', 'meta_keywords' => 'Epigenetics, DNA Methylation,5-hmC monoclonal antibody,hMeDIP,Bisulfite conversion,Methylated DNA immunoprecipitation', 'meta_description' => 'Complete, optimized solutions for analyzing DNA methylation manually or on our automated system.', 'meta_title' => 'DNA Methylation - Bisulfite sequencing - Epigenetics | Diagenode', 'modified' => '2019-03-25 10:07:27', 'created' => '2015-05-03 13:47:53', 'ProductsApplication' => array( [maximum depth reached] ) ) ), 'Category' => array( (int) 0 => array( 'id' => '87', 'position' => '1', 'parent_id' => '86', 'name' => 'PCR enzymes and Mixes', 'description' => '', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'PCR-enzymes-and-mixes', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and RT-PCR Mastermix ', 'meta_description' => 'Diagenode offers DNA polymerase and RT-PCR Mastermix ', 'meta_title' => 'PCR enzymes and RT-PCR Mastermix | Diagenode ', 'modified' => '2016-01-21 15:01:47', 'created' => '2015-09-16 23:10:49', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 1 => array( 'id' => '56', 'position' => '6', 'parent_id' => '12', 'name' => 'PCR enzymes and nucleotides', 'description' => '', 'no_promo' => false, 'in_menu' => false, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'pcr-enzymes-and-nucleotides', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and nucleotides, methylated DNA ,DNA methylation', 'meta_description' => 'Diagenode's PCR enzymes and nucleotides Optimized for amplification of methylated DNA ', 'meta_title' => 'PCR enzymes and nucleotides for DNA methylation | Diagenode', 'modified' => '2019-07-03 10:42:59', 'created' => '2015-07-08 09:57:30', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 2 => array( 'id' => '54', 'position' => '1', 'parent_id' => '12', 'name' => 'Bisulfite conversion', 'description' => '<div class="row"> <div class="small-12 medium-8 large-8 columns"><br /> <p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p> </div> <div class="small-12 medium-4 large-4 columns"><center> <script>// <![CDATA[ var date = new Date(); var heure = date.getHours(); var jour = date.getDay(); var semaine = Math.floor(date.getDate() / 7) + 1; if (jour === 2 && ( (heure >= 9 && heure < 9.5) || (heure >= 18 && heure < 18.5) )) { document.write('<a href="https://us02web.zoom.us/j/85467619762"><img src="https://www.diagenode.com/img/epicafe-ON.gif"></a>'); } else { document.write('<a href="https://go.diagenode.com/l/928883/2023-04-26/3kq1v"><img src="https://www.diagenode.com/img/epicafe-OFF.png"></a>'); } // ]]></script> </center></div> </div> <h2>How it works</h2> <p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p> <p style="text-align: center;"><span></span></p> <p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p> <h2>Advantages</h2> <ul class="nobullet" style="font-size: 19px;"> <li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li> <li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li> <li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li> </ul> <h2>Downstream analysis techniques</h2> <ul class="square"> <li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li> <li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li> </ul> <p></p>', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'bisulfite-conversion', 'cookies_tag_id' => null, 'meta_keywords' => 'Bisulfite conversion,bisulfite sequencing,DNA methylation,Epigenetics ,next-generation sequencing', 'meta_description' => 'Bisulfitre conversion is the gold standard method for DNA methylation studies at a single base pair resolution. 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Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p> </div> <div class="small-12 medium-4 large-4 columns text-center"><a href="../landing-pages/dna-methylation-grant-applications"><img src="https://www.diagenode.com/img/banners/banner-dna-grant.png" alt="" /></a></div> <div style="text-align: justify;" class="small-12 medium-12 large-12 columns"> <p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p> </div> <div class="small-12 medium-12 large-12 columns"><br /><br /> <ul class="accordion" data-accordion=""> <li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a> <div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base. <p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p> </div> </li> </ul> <br /> <h2>Main DNA methylation technologies</h2> <p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p> <div class="row"> <ol> <li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li> </ol> <p><span style="font-weight: 400;"> </span></p> <div class="row"> <table> <thead> <tr> <th></th> <th>Description</th> <th width="350">Features</th> </tr> </thead> <tbody> <tr> <td><strong>Bisulfite conversion</strong></td> <td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><b>Methylated DNA enrichment</b></td> <td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><strong>Restriction enzyme-based digestion</strong></td> <td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li> </ul> </td> </tr> </tbody> </table> </div> </div> <div class="row"></div> </div> </div> <div class="large-12 columns"></div> </div>', 'in_footer' => true, 'in_menu' => true, 'online' => true, 'tabular' => true, 'slug' => 'epigenetics-dna-methylation', 'meta_keywords' => 'Epigenetics, DNA Methylation,5-hmC monoclonal antibody,hMeDIP,Bisulfite conversion,Methylated DNA immunoprecipitation', 'meta_description' => 'Complete, optimized solutions for analyzing DNA methylation manually or on our automated system.', 'meta_title' => 'DNA Methylation - Bisulfite sequencing - Epigenetics | Diagenode', 'modified' => '2019-03-25 10:07:27', 'created' => '2015-05-03 13:47:53', 'ProductsApplication' => array( [maximum depth reached] ) ) ), 'Category' => array( (int) 0 => array( 'id' => '87', 'position' => '1', 'parent_id' => '86', 'name' => 'PCR enzymes and Mixes', 'description' => '', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'PCR-enzymes-and-mixes', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and RT-PCR Mastermix ', 'meta_description' => 'Diagenode offers DNA polymerase and RT-PCR Mastermix ', 'meta_title' => 'PCR enzymes and RT-PCR Mastermix | Diagenode ', 'modified' => '2016-01-21 15:01:47', 'created' => '2015-09-16 23:10:49', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 1 => array( 'id' => '56', 'position' => '6', 'parent_id' => '12', 'name' => 'PCR enzymes and nucleotides', 'description' => '', 'no_promo' => false, 'in_menu' => false, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'pcr-enzymes-and-nucleotides', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and nucleotides, methylated DNA ,DNA methylation', 'meta_description' => 'Diagenode's PCR enzymes and nucleotides Optimized for amplification of methylated DNA ', 'meta_title' => 'PCR enzymes and nucleotides for DNA methylation | Diagenode', 'modified' => '2019-07-03 10:42:59', 'created' => '2015-07-08 09:57:30', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 2 => array( 'id' => '54', 'position' => '1', 'parent_id' => '12', 'name' => 'Bisulfite conversion', 'description' => '<div class="row"> <div class="small-12 medium-8 large-8 columns"><br /> <p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p> </div> <div class="small-12 medium-4 large-4 columns"><center> <script>// <![CDATA[ var date = new Date(); var heure = date.getHours(); var jour = date.getDay(); var semaine = Math.floor(date.getDate() / 7) + 1; if (jour === 2 && ( (heure >= 9 && heure < 9.5) || (heure >= 18 && heure < 18.5) )) { document.write('<a href="https://us02web.zoom.us/j/85467619762"><img src="https://www.diagenode.com/img/epicafe-ON.gif"></a>'); } else { document.write('<a href="https://go.diagenode.com/l/928883/2023-04-26/3kq1v"><img src="https://www.diagenode.com/img/epicafe-OFF.png"></a>'); } // ]]></script> </center></div> </div> <h2>How it works</h2> <p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p> <p style="text-align: center;"><span></span></p> <p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p> <h2>Advantages</h2> <ul class="nobullet" style="font-size: 19px;"> <li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li> <li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li> <li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li> </ul> <h2>Downstream analysis techniques</h2> <ul class="square"> <li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li> <li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li> </ul> <p></p>', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'bisulfite-conversion', 'cookies_tag_id' => null, 'meta_keywords' => 'Bisulfite conversion,bisulfite sequencing,DNA methylation,Epigenetics ,next-generation sequencing', 'meta_description' => 'Bisulfitre conversion is the gold standard method for DNA methylation studies at a single base pair resolution. 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Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p> </div> <div class="small-12 medium-4 large-4 columns text-center"><a href="../landing-pages/dna-methylation-grant-applications"><img src="https://www.diagenode.com/img/banners/banner-dna-grant.png" alt="" /></a></div> <div style="text-align: justify;" class="small-12 medium-12 large-12 columns"> <p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p> </div> <div class="small-12 medium-12 large-12 columns"><br /><br /> <ul class="accordion" data-accordion=""> <li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a> <div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base. <p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p> </div> </li> </ul> <br /> <h2>Main DNA methylation technologies</h2> <p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p> <div class="row"> <ol> <li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li> </ol> <p><span style="font-weight: 400;"> </span></p> <div class="row"> <table> <thead> <tr> <th></th> <th>Description</th> <th width="350">Features</th> </tr> </thead> <tbody> <tr> <td><strong>Bisulfite conversion</strong></td> <td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><b>Methylated DNA enrichment</b></td> <td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><strong>Restriction enzyme-based digestion</strong></td> <td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li> </ul> </td> </tr> </tbody> </table> </div> </div> <div class="row"></div> </div> </div> <div class="large-12 columns"></div> </div>', 'in_footer' => true, 'in_menu' => true, 'online' => true, 'tabular' => true, 'slug' => 'epigenetics-dna-methylation', 'meta_keywords' => 'Epigenetics, DNA Methylation,5-hmC monoclonal antibody,hMeDIP,Bisulfite conversion,Methylated DNA immunoprecipitation', 'meta_description' => 'Complete, optimized solutions for analyzing DNA methylation manually or on our automated system.', 'meta_title' => 'DNA Methylation - Bisulfite sequencing - Epigenetics | Diagenode', 'modified' => '2019-03-25 10:07:27', 'created' => '2015-05-03 13:47:53', 'ProductsApplication' => array( [maximum depth reached] ) ) ), 'Category' => array( (int) 0 => array( 'id' => '87', 'position' => '1', 'parent_id' => '86', 'name' => 'PCR enzymes and Mixes', 'description' => '', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'PCR-enzymes-and-mixes', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and RT-PCR Mastermix ', 'meta_description' => 'Diagenode offers DNA polymerase and RT-PCR Mastermix ', 'meta_title' => 'PCR enzymes and RT-PCR Mastermix | Diagenode ', 'modified' => '2016-01-21 15:01:47', 'created' => '2015-09-16 23:10:49', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 1 => array( 'id' => '56', 'position' => '6', 'parent_id' => '12', 'name' => 'PCR enzymes and nucleotides', 'description' => '', 'no_promo' => false, 'in_menu' => false, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'pcr-enzymes-and-nucleotides', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and nucleotides, methylated DNA ,DNA methylation', 'meta_description' => 'Diagenode's PCR enzymes and nucleotides Optimized for amplification of methylated DNA ', 'meta_title' => 'PCR enzymes and nucleotides for DNA methylation | Diagenode', 'modified' => '2019-07-03 10:42:59', 'created' => '2015-07-08 09:57:30', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 2 => array( 'id' => '54', 'position' => '1', 'parent_id' => '12', 'name' => 'Bisulfite conversion', 'description' => '<div class="row"> <div class="small-12 medium-8 large-8 columns"><br /> <p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p> </div> <div class="small-12 medium-4 large-4 columns"><center> <script>// <![CDATA[ var date = new Date(); var heure = date.getHours(); var jour = date.getDay(); var semaine = Math.floor(date.getDate() / 7) + 1; if (jour === 2 && ( (heure >= 9 && heure < 9.5) || (heure >= 18 && heure < 18.5) )) { document.write('<a href="https://us02web.zoom.us/j/85467619762"><img src="https://www.diagenode.com/img/epicafe-ON.gif"></a>'); } else { document.write('<a href="https://go.diagenode.com/l/928883/2023-04-26/3kq1v"><img src="https://www.diagenode.com/img/epicafe-OFF.png"></a>'); } // ]]></script> </center></div> </div> <h2>How it works</h2> <p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p> <p style="text-align: center;"><span></span></p> <p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p> <h2>Advantages</h2> <ul class="nobullet" style="font-size: 19px;"> <li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li> <li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li> <li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li> </ul> <h2>Downstream analysis techniques</h2> <ul class="square"> <li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li> <li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li> </ul> <p></p>', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'bisulfite-conversion', 'cookies_tag_id' => null, 'meta_keywords' => 'Bisulfite conversion,bisulfite sequencing,DNA methylation,Epigenetics ,next-generation sequencing', 'meta_description' => 'Bisulfitre conversion is the gold standard method for DNA methylation studies at a single base pair resolution. 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Diagenode offers sonication instruments, reagent kits, high quality antibodies, and high-throughput automation capability to address all of your specific DNA methylation analysis requirements.</p> </div> <div class="small-12 medium-4 large-4 columns text-center"><a href="../landing-pages/dna-methylation-grant-applications"><img src="https://www.diagenode.com/img/banners/banner-dna-grant.png" alt="" /></a></div> <div style="text-align: justify;" class="small-12 medium-12 large-12 columns"> <p>DNA methylation was the first discovered epigenetic mark and is the most widely studied topic in epigenetics. <em>In vivo</em>, DNA is methylated following DNA replication and is involved in a number of biological processes including the regulation of imprinted genes, X chromosome inactivation. and tumor suppressor gene silencing in cancer cells. Methylation often occurs in cytosine-guanine rich regions of DNA (CpG islands), which are commonly upstream of promoter regions.</p> </div> <div class="small-12 medium-12 large-12 columns"><br /><br /> <ul class="accordion" data-accordion=""> <li class="accordion-navigation"><a href="#dnamethyl"><i class="fa fa-caret-right"></i> Learn more</a> <div id="dnamethyl" class="content">5-methylcytosine (5-mC) has been known for a long time as the only modification of DNA for epigenetic regulation. In 2009, however, Kriaucionis discovered a second methylated cytosine, 5-hydroxymethylcytosine (5-hmC). The so-called 6th base, is generated by enzymatic conversion of 5-methylcytosine (5-mC) into 5-hydroxymethylcytosine by the TET family of oxygenases. Early reports suggested that 5-hmC may represent an intermediate of active demethylation in a new pathway which demethylates DNA, converting 5-mC to cytosine. Recent evidence fuel this hypothesis suggesting that further oxidation of the hydroxymethyl group leads to a formyl or carboxyl group followed by either deformylation or decarboxylation. The formyl and carboxyl groups of 5-formylcytosine (5-fC) and 5-carboxylcytosine (5-caC) could be enzymatically removed without excision of the base. <p class="text-center"><img src="https://www.diagenode.com/img/categories/kits_dna/dna_methylation_variants.jpg" /></p> </div> </li> </ul> <br /> <h2>Main DNA methylation technologies</h2> <p style="text-align: justify;">Overview of the <span style="font-weight: 400;">three main approaches for studying DNA methylation.</span></p> <div class="row"> <ol> <li style="font-weight: 400;"><span style="font-weight: 400;">Chemical modification with bisulfite – Bisulfite conversion</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Enrichment of methylated DNA (including MeDIP and MBD)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Treatment with methylation-sensitive or dependent restriction enzymes</span></li> </ol> <p><span style="font-weight: 400;"> </span></p> <div class="row"> <table> <thead> <tr> <th></th> <th>Description</th> <th width="350">Features</th> </tr> </thead> <tbody> <tr> <td><strong>Bisulfite conversion</strong></td> <td><span style="font-weight: 400;">Chemical conversion of unmethylated cytosine to uracil. Methylated cytosines are protected from this conversion allowing to determine DNA methylation at single nucleotide resolution.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Single nucleotide resolution</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Quantitative analysis - methylation rate (%)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Gold standard and well studied</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><b>Methylated DNA enrichment</b></td> <td><span style="font-weight: 400;">(Hydroxy-)Methylated DNA is enriched by using specific antibodies (hMeDIP or MeDIP) or proteins (MBD) that specifically bind methylated CpG sites in fragmented genomic DNA.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Resolution depends on the fragment size of the enriched methylated DNA (300 bp)</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Qualitative analysis</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Compatible with automation</span></li> </ul> </td> </tr> <tr> <td><strong>Restriction enzyme-based digestion</strong></td> <td><span style="font-weight: 400;">Use of (hydroxy)methylation-sensitive or (hydroxy)methylation-dependent restriction enzymes for DNA methylation analysis at specific sites.</span></td> <td> <ul style="list-style-type: circle;"> <li style="font-weight: 400;"><span style="font-weight: 400;">Determination of methylation status is limited by the enzyme recognition site</span></li> <li style="font-weight: 400;"><span style="font-weight: 400;">Easy to use</span></li> </ul> </td> </tr> </tbody> </table> </div> </div> <div class="row"></div> </div> </div> <div class="large-12 columns"></div> </div>', 'in_footer' => true, 'in_menu' => true, 'online' => true, 'tabular' => true, 'slug' => 'epigenetics-dna-methylation', 'meta_keywords' => 'Epigenetics, DNA Methylation,5-hmC monoclonal antibody,hMeDIP,Bisulfite conversion,Methylated DNA immunoprecipitation', 'meta_description' => 'Complete, optimized solutions for analyzing DNA methylation manually or on our automated system.', 'meta_title' => 'DNA Methylation - Bisulfite sequencing - Epigenetics | Diagenode', 'modified' => '2019-03-25 10:07:27', 'created' => '2015-05-03 13:47:53', 'ProductsApplication' => array( [maximum depth reached] ) ) ), 'Category' => array( (int) 0 => array( 'id' => '87', 'position' => '1', 'parent_id' => '86', 'name' => 'PCR enzymes and Mixes', 'description' => '', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'PCR-enzymes-and-mixes', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and RT-PCR Mastermix ', 'meta_description' => 'Diagenode offers DNA polymerase and RT-PCR Mastermix ', 'meta_title' => 'PCR enzymes and RT-PCR Mastermix | Diagenode ', 'modified' => '2016-01-21 15:01:47', 'created' => '2015-09-16 23:10:49', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 1 => array( 'id' => '56', 'position' => '6', 'parent_id' => '12', 'name' => 'PCR enzymes and nucleotides', 'description' => '', 'no_promo' => false, 'in_menu' => false, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'pcr-enzymes-and-nucleotides', 'cookies_tag_id' => null, 'meta_keywords' => 'PCR enzymes and nucleotides, methylated DNA ,DNA methylation', 'meta_description' => 'Diagenode's PCR enzymes and nucleotides Optimized for amplification of methylated DNA ', 'meta_title' => 'PCR enzymes and nucleotides for DNA methylation | Diagenode', 'modified' => '2019-07-03 10:42:59', 'created' => '2015-07-08 09:57:30', 'ProductsCategory' => array( [maximum depth reached] ), 'CookiesTag' => array([maximum depth reached]) ), (int) 2 => array( 'id' => '54', 'position' => '1', 'parent_id' => '12', 'name' => 'Bisulfite conversion', 'description' => '<div class="row"> <div class="small-12 medium-8 large-8 columns"><br /> <p>Sodium bisulfite conversion of genomic DNA is the most commonly used method for DNA methylation studies providing <strong>single nucleotide resolution</strong>. It enables <span>to differentiate and detect unmethylated versus methylated cytosines. This procedure can then be followed either by <strong>PCR amplification</strong> or <strong>next generation sequencing</strong> to reveal the methylation status of every cytosine in gene specific amplification or whole genome amplification.</span></p> </div> <div class="small-12 medium-4 large-4 columns"><center> <script>// <![CDATA[ var date = new Date(); var heure = date.getHours(); var jour = date.getDay(); var semaine = Math.floor(date.getDate() / 7) + 1; if (jour === 2 && ( (heure >= 9 && heure < 9.5) || (heure >= 18 && heure < 18.5) )) { document.write('<a href="https://us02web.zoom.us/j/85467619762"><img src="https://www.diagenode.com/img/epicafe-ON.gif"></a>'); } else { document.write('<a href="https://go.diagenode.com/l/928883/2023-04-26/3kq1v"><img src="https://www.diagenode.com/img/epicafe-OFF.png"></a>'); } // ]]></script> </center></div> </div> <h2>How it works</h2> <p style="text-align: left;">Treatment of DNA with sodium bisulfite converts unmethylated cytosine to uracil, while methylated cytosines remain unchanged. <span>The DNA is then amplified by PCR where the uracils are converted to thymines. </span></p> <p style="text-align: center;"><span></span></p> <p><img src="https://www.diagenode.com/img/categories/bisulfite-conversion/bisulfite-conversion-acgautac.png" style="display: block; margin-left: auto; margin-right: auto;" /></p> <h2>Advantages</h2> <ul class="nobullet" style="font-size: 19px;"> <li><i class="fa fa-arrow-circle-right"></i><strong> </strong><strong>Single nucleotide</strong> resolution</li> <li><i class="fa fa-arrow-circle-right"></i><strong> Gene-specific </strong>and <strong>genome-wide</strong><span> analyses</span></li> <li><i class="fa fa-arrow-circle-right"></i><strong> NGS</strong><span> </span>compatible</li> </ul> <h2>Downstream analysis techniques</h2> <ul class="square"> <li>Reduced Representation Bisulfite Sequencing (RRBS) with our <a href="https://www.diagenode.com/en/p/premium-rrbs-kit-V2-x24">Premium RRBS Kit V2</a></li> <li>Bisulfite conversion with our <a href="https://www.diagenode.com/en/p/premium-bisulfite-kit-50-rxns">Premium Bisulfite Kit</a> followed by qPCR, Sanger, Pyrosequencing</li> </ul> <p></p>', 'no_promo' => false, 'in_menu' => true, 'online' => true, 'tabular' => true, 'hide' => false, 'all_format' => false, 'is_antibody' => false, 'slug' => 'bisulfite-conversion', 'cookies_tag_id' => null, 'meta_keywords' => 'Bisulfite conversion,bisulfite sequencing,DNA methylation,Epigenetics ,next-generation sequencing', 'meta_description' => 'Bisulfitre conversion is the gold standard method for DNA methylation studies at a single base pair resolution. 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