DNA quality and purity for pyrosequencing experiments?

31 July 2018 1 10K Report

Is there a 260/280 ratio cutoff for DNA that is to be used for bisulfite conversion and pyrosequencing? How pure does the DNA have to be?

Abhijeet Singh

https://www.illumina.com/documents/products/appnotes/appnote-methylseq-wgbs.pdf

As per Illumina, there is no specific requirement for the DNA quality with respect to the bisulfite sequencing. Thus, DNA quality of A260/A280 should be 1.8-2.0 (which is the general quality requirement for DNA). Also, if possible, use qubit for the quantification, as nanodrop can give false quantification of the DNA.

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