Differential Display

Hank Seifert h-seifert at nwu.edu
Sat May 21 17:59:29 EST 1994


In article <199405210231.TAA19524 at net.bio.net> IFOO%UKANVM.bitnet at VM42.CSO.UIUC.EDU (Ivy Foo) writes:
>I have been trying differential display for the last few months.  My
>sequencing gels look great and I am able to isolate and reamplify the
>'differential'bands.  However, I am having all sorts of problems with
>comfirming the bands on Northerns.  I have been making random primed
>probes with the reamplified bands and hybridizing to Northerns as
>suggested by GenHunter Co., however, no signal ever appears on the
>autorads! From what I've been reading on the Net it would appear that
>just about everyone doing DD is stuck on this step....any comments
>or ideas as to how we can get around this without undergoing massive
>cloning and screening??  Thanks!!
>
>-------------------------------------------------------------------------
>Ivy Foo
>Department of Biochemistry and Molecular Biology
>University of Kansas Medical Center
>Kansas City, KS 66202
>(913)58-7425
>BITNET : IFOO at UKANVM                         ››››/
>INTERNET : IFOO at UKANVM.CC.UKANS.EDU          (@ @)
>------------------------------------------OOO (_) OOO--------------------
Dear Ivy,
I haven't done the technique but I have seen that some people have better 
results using RT-PCR with their primers then Northerns to detect 
the mRNA.  I'm 
getting the feeling that DD often produces rare messages that 
are difficult to detect in Northerns (and then difficult to 
clone from a cDNA library!).  Don't forget to do the no RT 
control.

I hope this helps

Hank Seifert
Northwestern University




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