RAPD Trouble!!!

[Roland Koelliker]

Dear netters
I'm using RAPD to detect genetic variability in festuca pratensis (meadow
fescue). All worked fine so far, after initial problems I found a set of 20
primers that produce nice polymorphic bands. All of a sudden these primers
stop to work. I tried everything (new polymerase, new buffer, mgcl, dntp,
and so on) but still I do not get the same nice results as before. Since it
once worked and I already have some nice pictures I don't want to play
around with the reaction conditions again. Does anybody have an idea what's
going on here? I'm quite desperate.

Thank your help

Roland



Roland Koelliker	                  
Swiss Fed Inst Technol               
Plant Sciences                              
ETH-Zentrum, LFW-C47         
CH-8092 Zurich                          
Tel ++41-1-632 4890
Fax ++41-1-632 1153
e-mail: koelliker at ipw.agrl.ethz.ch