CpG Islands
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. EMD Millipore’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.
CpG 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. EMD Millipore’s CpGenome Turbo Bisulfite Modification kit provides a unique set of reagents for fast and effective bisulfate modification with as little as 1 ng of DNA, and additional spin columns facilitate easy isolation of bisulfate-modified DNA. With the CpGenome Turbo Bisulfite Modification kit this assay can be completed in as little as 90 minutes.
Methyl binding domain pull down
The Methylbinding Domain Pull Down assay employs a high-affinity recombinant glutathione-S-transferase (GST) tag that has been fused with the methylbinding domain (MBD) of a methyl CpG binding protein, to selectively bind methylated DNA in cell or tissue lysates. EMD Millipore’s CpG MethylQuest DNA isolation kit features a high affinity GST-MBD coupled to magnetic beads for easy and effective enrichment of methylated DNA at a specific locus or within the entire genome.
Methylated DNA IP
EMD Millipore provides a range of high quality antibodies and bead technology that can selectively bind methylcytosine residues in cell or tissue samples. Enriched methyl-DNA can be amplified and quantified by qPCR, and is also suitable for genome-wide microarrays or sequencing.
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. 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 methylation patterns using as little as 1 ng of DNA.