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Bisulfite Conversion Efficiency - (Apr/28/2010 )

Does anyone know what is the best way to determine bisulfite conversion efficiency? i.e. would MSP or bisulfite sequencing be used for this?
Also I am trying to optimize bisulfite conversion efficiency of FFPE samples....does anyone have experience with this?

-Boba-

this is difficult to determine properly because the primers you use are designed to bias towards fully converted template.

Efficiency is then measured by cloning and sequencning methods where non-CpG C's sequenced are counted across all the clones sequenced.

Note this is only conversion efficiency of the region you are interested in and PCR amplified, and not necessarily a measure of efficiency across the whole genome. There are some sequence contexts where conversion could be inhibited by GC-content and secondary structures.

-methylnick-