help for oligo hybridization

Mic Chaudoir mic at nwu.edu
Mon Oct 31 17:07:21 EST 1994


In article <38nigg$2u3 at nntp.Stanford.EDU>, minmin at leland.Stanford.EDU
(Limin Li) wrote:

> I have been trying different conditions for oligo(25-40 mers)
hybridization.It \
> seems I always get strong background or nothing(of course I tried
different tem\
> peratures and washing conditions). I am using Amersham's filter. I
checked the \
> oligos on computer, they all ok.
> Any suggestions would be greatly appreciated.
> Thanks in advance.
> 
> Limin Li
> Dept. of Genetics
> Stanford U. Sch. of Medicine
> Stanford, CA 943050-5120

The RebBook has a protocol which I have used.  It works fine for me. 
However, I have not used it for a real southerns, only detection of T3/T&
oligos in a piece of phage DNA.  Thus, I had ng levels of DNA on the
filter, not pg.  This probably helps a lot.  Additionally, we use the
"GeneScreen" filter, which always seems tow rok really well for us.

-- 
mic
mic at nwu.edu

"DOS is proof that PC users can take a joke"



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