PCR'd DNA vs RNA probes for northerns

Vivian Miao vmiao at oregon.uoregon.edu
Sat Oct 30 14:26:38 EST 1993


In article <2au7g6$ojo at usenet.INS.CWRU.Edu>, Harry Menegay wrote:
> 

> 	We are currently making RNA probes to probe northern blots, however we are
> considering going to DNA probes made from single pimer PCR.  Has anyone tried one
> versus the other? Is background better or worse? Is the signal decreased?

Hi Harry,  I don't know if this addresses your question about background,
but my "normal" probes for Northerns are made by the random hex method off
a gel isolated fragment or off a PCR product, so presumably both strands
are there.  I know half the probe is going to waste, but the signal is
fine, and with respect to your concern - there isn't any background
problem.  I work with Neurospora crassa, so maybe the small genome (43
megabases) has something to do with why I can get away with this.  

I also have a protocol for making DNA probes by "single primer PCR" (linear
amplification) which I'll be glad to post or email if it is of interest.



More information about the Methods mailing list