PCR'd DNA vs RNA probes for northerns
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.
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