PCR from BAC - (Jul/20/2004 )
I am trying to PCR approx 400bp fragments from a BAC clone with no success. I have successfully used these primer sets from genomic DNA in a related animal. I have no sequence information for this clone but we are trying to amplify fragments that are well conserved in other species. Are there any special procedures in PCR'ing from BAC's. Should I try to cdegest the BAC before PCR. I am without a clue. Thank you for help.
How much amount of BAC DNA is used per reaction? I generally don't use more than 50 ng per 20 ul reaction. You may also want to try the flanking BACs in the tiling path.