DNA methylation - (Oct/28/2004 )
Hi, I am having troubles with PCR after the DNA sulphitation (to study) DNA methylation. I think it is because of the low recuperation after the sulphite reaction.
Does somebody have experience with that?
I don't understand what you mean by "recuperation".
Many things contribute to failed PCR after DNA modification, such as unsuccessful modification, loss of DNA, poor primer design, not optimized PCR conditions... Please make your question clear and provide more details.
Sorry, I think it is loss of DNA during the reaction.
How did you purify your DNA after modification? Did you use a kit for the modification? DNA loss during purification is a problem but not the only problem for PCR failure. Try use some controls if you can get it such as good modified DNA or known working primers.