Methylation-specific PCR (MSP) analysis of DNA methylation patterns in CpG islands (CAT#: STEM-MB-0212-WXH)

Methylation-specific PCR (MSP) is a PCR-based method for the analysis of methylation patterns in CpG islands. To perform the MSP assay, DNA is purified and undergoes modification by using sodium bisulfate, in a sulphonation process that converts all unmethylated cytosine residues to uracil. Methylated cytosine, in which a methyl group is attached to carbon 5, cannot be converted since the sulphonation reaction cannot occur.
For a MSP experiment, two pairs of primers are needed. One pair is specific for methylated DNA (M) and the other is specific for unmethylated DNA (U). For discrimination of methylated and unmethylated DNA, one or more CpG sites are included in each primer (or at least one of the pair) sequence.
PCR reactions are performed using each primer pair, M and U primer pair.
Successful amplification from the M primer pair is indicative of methylated DNA and PCR product by U primer pair is reflective of unmethylated DNA.

• Detect genes or sequences with DNA methylation.
• Early tumor diagnosis, disease course monitoring and efficacy assessment.
• Study of genetic blotting, embryonic development.

1.Sample preparation
2.DNA isolation
3.DNA modification
4.Primer design
5.Methylation-specific PCR

Two pairs of primers are needed: One pair is specific for methylated DNA (M) and the other is specific for unmethylated DNA (U).