Abstract
The methylation profile of circulating cell-free DNA (cfDNA) in blood can be exploited to detect and diagnose cancer and other tissue pathologies and is therefore of great diagnostic interest. There is an urgent need for a cost-effective genome-wide methylation profiling method that is simple, robust and automatable and that works on highly fragmented cfDNA. We report on a novel sample preparation method for reduced representation bisulfite sequencing (RRBS), rigorously designed and customized for minute amounts of highly fragmented DNA. Our method works in particular on cfDNA from blood plasma. It is a performant and cost-effective methodology (termed cf-RRBS) which enables clinical cfDNA epigenomics studies.
Footnotes
The concentrations of the individual dNTPs was corrected for the dNTP-mix used during the Klenow(exo-) reaction of the cf-RRBS protocol. 0.25 uL of dNTP mix (4 mM dATP, 400 uM dCTP and 400 uM dGTP)