PCR from parraffin embeded samples - (Jan/02/2002 )
Anyone can give me a working protocol or suggestions on PCR amplifying DNA extracted from parraffin embeded sample? It is difficult to amplify relatively large fragment of DNA from parraffin sample. I thought the DNA extraction from parraffin sample is crutial for a successful PCR. My question is: 1. what is the best extraction protocol that can avoid fragmentation of the DNA during extraction? or it is all depend on the parraffin sample itself? I know ambion has kit for RNA extraction from parraffin sample but don't know if there is kit for DNA. 2. Does dilution of the DNA template helps? Maybe something in DNA which come from parraffin extraction inhibited the activity of DNA polymerase? Thanks for any suggestions.
(Edited by huangz at 3:31 am on Jan. 2, 2001)
what's your expected pcr size? Usually, around 400bp should be no problem. In my hand, any extraction method works, such as qiagen tissue kit, traditional phenol-chloroform extraction.
This URL has some useful information about pcr DNA from paraffin block