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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-