[bionet.molbio.methds-reagnts] Oligonucleotide probing

LAFLAMME@MAINE.BITNET (10/26/90)

Does anyone out there in netland have any experience in the use of
oligonucleotide probes to screen total DNA in northern or slot-blots?
I have been using 20 mers to pick out individual members of a family
of plant genes.  It seems that the method is not sensitive enough to
detect these sequences in a sample containing a HUGE excess of
non-hybridizing species.  If anyone could pass along a helpful hint
I would be very happy.  I would prefer not to isolate poly A RNA due
to the extra hassle and the possibility of skewing the final results.

mkw6d@krebs.acc.Virginia.EDU (Martyn K. White) (10/30/90)

In article <90298.171656LAFLAMME@MAINE.BITNET> LAFLAMME@MAINE.BITNET writes:
>Does anyone out there in netland have any experience in the use of
>oligonucleotide probes to screen total DNA in northern or slot-blots?
>I have been using 20 mers to pick out individual members of a family
>of plant genes.  It seems that the method is not sensitive enough to
(stuff deleted)

Have you thought of PCR.

	Martyn K. White, Box 441, U.Va, Cville VA 22908
	     		(804) 924 8710	
                 mkw6d@krebs.acc.Virginia.EDU
		      mkw6d@Virginia.EDU

IBT329%DJUKFA11@PUCC.PRINCETON.EDU (11/05/90)

It might be that asymmetric PCR is an answer:
i.e.
    Primer A = 50 - 100 pmol
    Primer B = 0.5 - 2  pmol

Bilanovic D.D