1 reply to this topic

[rajgene]

HI GUYS;
i designed a primer for a promotor region of fcerg1 gene to study its snps effects via reporter gene study.
i did a epcr of the primers from ncbi and it matched perfectly to the corresponding locus, but after i added a restriction sites to the forward and reverse primer mul I and Xho I respecitveöy for PCR cloning in my PGL2 basic vector, the primers dont score a epcr hit.
is this a serious problem.... will it affect the actual amplication of my product during real PCR.
please explain me how to proceed?
thxx
raj

[pcrman]

No, it won't. Because you have added some bases to the 5' end of both primers, the program can no longer find a match for the new sequences. So just go ahead with your PCR.