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How can I know complete bisulfite conversion - (Jun/28/2005 )

as titles, thanks a lot.

Also, there is always mention the low efficiency of the bisulfite sequencing primer. How can I perform trouble shooting?



As Nick said in your another post, you can cut any non-CPG sites with restriction enzyme. For example, the original sequence is GGCTAGCG, after modification and PCR, the sequence should be GGTTAGC*G (* unknown). If an enzyme recognizes GGCTAG and your DNA is fully converted, the enzyme should no longer cut that site.

You can also sequence your BSP product to see if all non-CpG 'C's have become 'T's.


also be sure to follow the methods of primer design as outlined in Warnecke et al 2002 Methods 27(2):101-7

Good luck!

Nick biggrin.gif