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CpG Islands & CpG Methylation

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Methylcytosine is relatively abundant in the human genome, accounting for approximately 1% of DNA bases. These methylated residues may have a profound effect on gene expression in eukaryotic cells. Several assays have been developed to evaluate the methylation status of DNA at either a single locus or within the entire genome. MilliporeSigma’s advanced line of kits, antibodies, and reagents simplifies these assays and allows for fast and effective enrichment of target DNA and further genome-wide analyses.

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(Click to enlarge.) Fastest Protocol Among Leading Suppliers. Comparison of time required for CpGenome™ Turbo bisulfite modification kit protocol versus bisulfite kit protocols from other major suppliers.

CpG Methylation Assays

Bisulfite modification
Bisulfite modification is widely used to distinguish between methylated and unmethylated cytosine residues. Sodium bisulfate is used to selectively deaminate cytosine residues to form uracil, whereas methylcytosine residues remain unchanged.

MilliporeSigma’s CpGenome Turbo Bisulfite Modification Kit is designed to simplify & streamline the bisulfite modification process to efficiently identify CpG Methylation sites. In just 90 minutes, go from DNA sample to bisulfite converted DNA ready for analysis.

MilliporeSigma’s CpGenome™ Direct Prep Bisulfite Modification Kit allows Bisulfite conversion directly from cultured cells, blood, fresh or fixed tissue.

Methyl Binding Domain Pull Down

Methylation-specific PCR (MSP) is an established technique for mapping and monitoring CpG methylation patterns in the CpG islands of genomic DNA. EMD Millipore’s CpGenome™ and CpG WIZ® systems allow sensitive detection of gene methylation using MSP of bisulfite-modified DNA. Rapid and simple, the method eliminates restriction digests and Southern blots. The CpGenome™ kit can detect changes in CpG methylation patterns using as little as 1 ng of DNA.

The CpGenome™ 5-hmC Quantitation Kit

Ideal for global and tissue-specific detection and quantitation of 5-hmC in genomic DNA, as well as high-throughput screening of activators and inhibitors of 5-hmC modification.
  • Compatible with genomic DNA from cultured cells or tissues 
  • Includes set of 5-hmC of standards for precise quantitation 
  • Streamlined, simple, automation-friendly ELISA-based protocol 
  • Sensitive colorimetric detection with a limit of quantitation down to 0.03% of input DNA.
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Simple ELISA-based Assay Procedure Reliable assay work flow allows for sensitive detection of 5-hmC in genomic DNA samples.

Click image to enlarge.
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Tissue-Specific Detection of 5-hmC Levels 100ng of genomic DNA from different tissues (CpGenome™ Mouse 5-mC and 5-hmC DNA Standards, Catalog No. S8004) was analyzed using CpGenome™ 5-hmC Quantitation kit. Data demonstrates reliable detection of differences in 5-hmC level of input samples consistent with those previously determined by LC/MS analysis (mouse brain 0.55%; mouse kidney 0.23%; mouse liver 0.11%; mouse thymus 0.03%)

Click image to enlarge.

Methylation Specific PCR Analysis

Methylation-specific PCR (MSP) is an established technique for mapping and monitoring methylation patterns in the CpG islands of genomic DNA. MilliporeSigma’s CpGenome™ and CpG WIZ® systems allow sensitive detection of gene methylation using MSP of bisulfite-modified DNA. Rapid and simple, the method eliminates restriction digests and Southern blots. The CpGenome™ kit can detect changes in methylation patterns using as little as 1 ng of DNA.