UBC primer PCR - (May/19/2008 )
Dear all,
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?
-genejockey-
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?
-Ambrósio-