To anyone on the net who knows:
I have been attempting to PCR a fragment on the Drosophila Simulans TRA gene with almost no luck. My primers are both 17 bases long and they span a segment of about 800 bases. I've tried many different annealing temperatures from 53 - 65 degrees Celsius, and have also tried different concentrations of genomic DNA, I've run 25 cycles, 30 cycles, and 35 cycles, and I still can't get the damn thing to work. Does anyone know anything I don't know about genomic fly DNA and the effective PCR of these DNA?
Please respond. My email address is: jalin at ucsd.edu Thanks.