How to measure the bisulfite concentration?

06 May 2024 1 9K Report

Does anyone know the exact BS concentration of the Zymo DNA methylation gold kit? or is there anyway to measure the BS concentration by titration? Thanks.

Mohammad Saber Zamani

According to the protocol of the EZ DNA Methylation-Gold Kit, the DNA recovery rate after bisulfite conversion is approximately 75%. On the other hand, due to the conversion of cytosine to uracil after bisulfite conversion, the two DNA strands no longer match, and the bisulfited DNA becomes single-stranded. Therefore, when using spectrophotometers, settings for single-stranded DNA must be applied.

There is also an older method to verify the presence of bisulfited DNA after bisulfite conversion. In this method, the bisulfited DNA is first subjected to electrophoresis and then placed on ice for 20 minutes. The result is observed using a Gel Doc device. The presence of a DNA smear indicates the presence of bisulfited DNA.

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