bisulphite conversion method comparison - (Dec/10/2006 )
Dear methylation experts,
What is the general verdict on bisulfite conversion technique- kits such as Epitect or Manual 16 hr. method of Herman et al? My positive universal methylated DNA and negative Normal Pbl give identical and correct result with both methods but in patient samples methylation bands are fewer with the kit.The faint methylated beands seen with the manual protocol disappear in the kit which is a 5 hr. conversion process as compared to 16hrs with the manual.We do MSP for methylation detection. This question has been posed before by me but I did not get enough answers to solve my dilemma. Please help me with this.Thanks in advance.
to solve your dilemma I would suggest that you use both methods and do bsp for your target sequence in some patient samples (as well as in your controlls). You can directly compare the efficacy of both methods there (complete conversion, etc) and decide afterwards, which one is best for your study.
Or am I wrong?