UBC primer PCR - (May/19/2008 )
I am trying the RAPD UBC primer sets and I was wondering what PCR conditions are most often used for these primers.
I tried this protocol last night:
1) 94C for 4 minutes
2) 94C for 1 minute
3) 37C for 1 minute
4) 72C for 2 minutes
5) Go to step 2) for 39 times
6) 72C for 7 minutes
I got NO bands at all, not even a smear.
What am I doing wrong? The DNA is good as I've used it for other kinds of PCR based markers but this is my first try with the UBC primers. Any suggestions?
Well, where did you get your primers from? Was it from an article or did you design them yourself?
Are you sure they work? Do you have any positive control?