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AnyGenes

Signaling Pathways title  Methylation Specific PCR

An innovative version of methylation-specific PCR, is MSqP (methylation specific quantitative PCR)  developed in our laboratory to detect and quantify the methylation of CpG islands from intergenic and intragenic regions of genomic DNA. The MSqP is a quantitative fluorescent-based real-time PCR method, it combines methylation specific priming with methylation specific fluorescent probing for genomic DNA converted by bisulfite reaction. As the conversion of cytosine to uracil generates non complementary DNA strands (U opposite to G), specific primer pair could be designed from either top or bottom DNA strand.

 

AnyGenes team has great experience in designing specific methylation primer assays to analyse methylation profile of one or several genes of your interest with one or several CpG islands (see figure below) 

  • For efficient analysis, we focus on CpG islands of less than 150 bp/region.
  • We use Actin as reference gene for the calculation of DCt= Ctref - Ctgene
  • For the relative methylation calculation, it is calculated as following:

           DDCt= DCtsample_test - DCtSample_control  

  • As controls we use genomic DNA of 0% methylation and 100% methylation.

As results you receive a report on the percentage methylation status of CpG islands of the genomic targeted region from your samples in Excel sheet with box plot, histogram illustrations, further analysis can be performed for p-value, and pathway analysis if needed…

  • Example: evaluation of methylation of MIR-129-2 gene in tumor and normal tissue of patients with gastric cancer.

 

If you need to explore the methylation profile of one or several genes, we can help you to perform this analysis from start to end, you have just to send us your:

  • Biological samples: biopsies, tissues, blood, FFPE…or genomic DNA
  • The name, accession number of your gene and the targeted region for methylation analysis

AnyGenes team has many years of experience in processing all kind of biological samples, we will perform the analysis for you in 2-4 weeks with a final report.