AnyGenes offers to analyze the methylation profile of your sample by pyrosequencing, in association with a bisulfite treatment.

In vertebrates, methylation of DNA is located on cysteine in CpG sites.
Bisulfite treatment of DNA will turn non methylated cysteine into uracil. After PCR amplification, non methylated cysteines will become thymidine. The remaining percent of cysteine will be representative of the methylation percentage in a given position of DNA.

Pyrosequencing is a sequencing method, where nucleotides are added 1 by 1. If a dNTP is used, it will release pyrophosphate which will be reused as ATP to produce fluorescence thanks to luciferase. So it is possible to know exactly which nucleotide was added at the moment. If the dNTPs are not used, they are degraded thanks to apyrase enzyme.

A sequence based detection technics Monitors the real time incorporation of nucleotides through enzymatic conversion of released pyrophosphate into a proportional light signal.

Pyrosequencing qPCR arrays
Pyrosequencing2 qPCR arrays

Thanks to our bioinformatics and data mining platform, AnyGenes scientists will help you to set up a complete service for your methylation studies.
For more informations, please contact us.