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Bisulfite Sequencing: establishment - (Feb/01/2008 )


I am trying to establish a pyrosequencing based methylation study in our lab. Unfortunatly, I got also signals for untreated genomic DNA with pyrosequencing. Any guess, how this can happen?

For the establishment, I tested several concetrations of bisulfite treated DNA and I also made mixtures of methylated and unmethylated DNA to measure the degree of methylation with pyrosequencing. Does anyone have some more clues how to establish the method properly?

Thanks for your help!



Can you tell us a bit more about the conversion and your primers?