DNA methylation profiling Gene specific profiling - A2780 TIMP2

The estimation of DNA methylation level heavily depends on the complete conversion of non-methylated DNA cytosines. It is crucial to ensure complete conversion of non-methylated cytosines in DNA. Therefore, it is important to incorporate controls for bisulfite reactions, as well as to pay attention to the appearance of cytosines in non-CpG sites after sequencing, which is an indicator of incomplete conversion.

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4 years ago

4 years ago by Jody Hancock Canada

How can I preserve leukocytes for DNA methylation profiling?

I would like to preserve leukocytes for future epigenetic analysis. How can I preserve them effectively in order to perform DNA methylation profiling at a later time?

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Protocol tips
According to the predicted CpG islands and published literature, we developed one pair of nested MSP primers that is listed in Supplemental Table . Genomic DNA of ovarian cancer cells was extracted using a DNA Isolation Kit (Tiangen Biotech, Beijing, China). Bisulfite modification of DNA was performed following the recommended procedures of the BisulFlash DNA Modification Kit (Epigentek, Farmingdale, NY,USA). Placenta DNA was modified with the CpG Methyltransferase (M.SssI, NewEngland Biolabs, Ipswitch, MA, USA) and used as a positive control.
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