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bisulfite treatment - in my hands bisulfite kit don't work well (Jan/22/2007 )

, i use 2 kit for bisulfite treatment and they are: EZ kit and qiagen kit
but I have a lot of problems...
when i use qiagen kit i have not amplicone but when i use EZ kit...i have amplicon but i have other problems...

i treat my genomic DNA and then I make a PCR using primers that have not modified...i have the amplicon and i clone in TOPOTA and then sequencing...
but when i analize the sequence of my gene promoter i see only few C convert in T...
and i see that cytosine also near other C, or near T and A don't convert...
how is it possible?
maybe my kits don't work well or my primers amplified only genomic DNA that is not modified completely...
have you observed this thing?

bye bye sad.gif


hi full-methyl,

I have been there before,

you said you used primers that were to unmodified DNA, this is the reason why you are seeing very low conversion rates.

Have a look at my pinned thread with regard to primer design.