5μg Purified DNA Methylation Analysis Bisulfite Sequencing BSP MSP Methylation Specific PCR

DNA methylation is a key epigenetic modification to cytosines, often in CpG dinucleotides. This modification has been frequently associated with gene silencing, but the precise role of DNA methylation in development and disease remains a mystery.

Bisulfite modification and sequencing method (BSP)

This method is reliable and precise. procedures:

1. Prepare genomic DNA of Sample
2. Bisulfite treatment (including purification and de-sulfosalicylic reaction) to convert unmethylated cytosine to uracil, while methylated cytosine remains unchanged.
3. Using methylation primer design software to design primers at the both ends of methylated regions.
4. Fragment amplification and PCR product recovery.
5. PCR products were sequenced directly or after cloned (quantitative analysis), analysis of each CpG island methylation.
6. Data Statistics. unmethylated DNA samples are used as a control.

Methylation specific PCR method (Methylation-Specific PCR, MS-PCR)

High sensitivity of this method is mainly used for qualitative research. Procedures:

1. Prepare genomic DNA of Sample
2. Bisulfite treatment (including the purification and de-sulfosalicylic reaction).
3. Using methylation primer design software design methylation specific primers (primer 1) or non-methylated primers in methylated regions (primer 2) 2 pairs.
4. For specific PCR amplification, only the fragments fully combinate with methylated or non-methylation specific primers can amplify the product.
5. Detection of MSP amplification, if DNA methylation for the treatment chain primers amplified fragments, it shows that the detected methylation sites exist; if used for the treatment of non-methylated DNA chain The primers amplified fragments, then the detected methylation sites does not exist.

Service content

Results include Lab report (experimental steps, including the BSP method statistical results of 5 clones), PCR electrophoresis photographs, sequencing trace files.