Reverse Northern
Ferland Louis H.
ferlandl at ERE.UMontreal.CA
Sat Sep 21 02:03:48 EST 1996
This is a message for Paul Hengen: we lose! (see below)
With apologies to those who haven't followed (or didn't care about) the
previous debate on the sense/antisense terminology.
On 21 Sep 1996, Gene Huh wrote:
[big snip]
>
> I actually dislike the term "Reverse Northern" the way I have seen it
> used, primarily because it is jargon-derived jargon. I mean, in many
^^^^^^^^^^^^^^^^^^^^^^(ouch!)
> cases, cDNA is used to probe DNA on dotblots. Seems like a Southern to me
> :). I wish there were a better term (e.g., Eberwein's lab also uses the
> term "expression profiling", which seems more accurate in that context).
>
>
> Incidentally, while I am getting off my soapbox <g>, I was wondering
> if anyone else out there has had luck with the technique Eberwein's lab
> describes. Briefly, a T7 RNA polymerase promoter-oligo dT primer was used
^^^
> to make ds-cDNA. T7 RNA polymerase was then used to generate aRNA
^^^^^^^^(the funniest thing is I *do* know what you mean!)
> (antisense RNA, same sequence as first strand cDNA). The process may be
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^(this one for Paul)
> repeated (ds-cDNA synthesis followed by T7 transcription) to reamplify the
> original starting mRNA population. Apparently, the degree of
> amplification can potentially be up to a few orders of magnitude for each
> cDNA/transcription cycle. Any comments?
>
> Apologizing for my semantic nitpicking and
^^^^^^^^^^^^^^^^^^^Gene! Huh? (Sorry for this...)
> Thanking you in advance,
>
> Dr. Gene Huh
> Department of Molecular and Cell Biology,
> University of California at Berkeley
> Berkeley, CA 94720-3200
> Email: gshuh at garnet.berkeley.edu
>
Dr. Louis H. Ferland
Centre de Recherche, Hotel-Dieu de Montreal
Dept de Nutrition, Universite de Montreal
Phone: (514) 843-2757 FAX: (514) 843-2719
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