Amplification of human genomic DNA - (Jan/05/2010 )
I am trying to amplify the ACTB gene in human cells. I design primers to amplify the genomic DNA but after sequencing my PCR product (and obviously based also on the size), I amplified DNA without introns. I tried to treat my genomic DNA with RNase but nothing changed. I am working with Hela and 293T cells. Could it be the problem ? Should I try to amplify it from primary cells ?
how did you design your primers. are you sure they are specific for that gene? NCBI has a primer designing software which enables you to design specific primers.
I designed new primers and it works !