PCR with FISH problems

[kvernick at helix.nih.gov]

> 
> Can anyone tell me why FISH works well with an original set of
> probes but when I PCR them, I get lots of non specific binding.
> 
> 				Very Frustrated :-[


Did you gel purify the PCR products? PCR generates HMW garbage which can give the same 
problem, nonspecific binding, when labelled and used as a probe for a S or N blot. Gel 
purification of the product away from LMW (primers) and HMW junk alleviates the problem. 
Hope this helps.

Ken