Weird Northern again!!!

Chunxin Wang cxwang at
Tue Aug 1 12:56:37 EST 2000

On 25 Jul 2000, Michael L. Sullivan wrote:

> How are you preparing the PCR fragments for labeling?  Are they straight
> out of a PCR reaction, cleaned up somehow, or have you cloned them into a
> vector and then using fragment prepared from that.  I was thinking that
> maybe if you are using dircetly out of the PCR reaction, perhaps you are
> labeling something that is relatively low abbundance, but hybridizes well
> to rRNA.  Since there is so much rRNA on your blot, those bands could get
> really hot and mask your real signal.  Could this be?
Initially I suspect my labeling kit might be contaminated (or the water),
so I bought a new one and made two probes again at the same time. Now one
probe worked but another one still hybridized to rRNAs. One conclusion was
made from the past results that all the probes made from PCR hybridized to
rRNA while the probes made from EST clones worked well.

So, now I totally agreed with you.



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