5' RACE

vilimf01 at mcrcr6.med.nyu.edu vilimf01 at mcrcr6.med.nyu.edu
Mon Mar 18 16:41:20 EST 1996


After doing RACE a number of times unsucessfully, I finally got
clontech's old amplifinder kit to work.  I did, however have to play
around with PCR additives to get one of them to work at all. 5-10%
DMSO or 5% formamide did wonders.  I don't particularly like the new
marathon kit from clontech because, in making second strand, some
length of message is usually lost.  I couldn't afford this as I only
had 10-13 bases of 5' untranslated.  This and most kits recommend use
of poly A RNA.  You may get lucky with clontech's amlpifinder ready
cDNA if they carry the source you are interested in and if your
message is abundant enough.  That may save you some time and effort.

Good luck			Usual disclaimers
							SVEN



In article <4h77iq$7dh at tribune.usask.ca>, bonhamp at duke.usask.ca (Peta C. Bonham-Smith) writes:
> I need to carry out some 5' RACE to locate the 5' end of a message of 
> interest - do I need Poly A+ RNA for this procedure?  If so what is the 
> recommended procedure these days - I really don't want to go back to 
> oligo-dT columns!!!  Secondly, which kit or procedure could you recommend 
> for the 5' RACE itself?
> 
> Please reply to
> bonhamp at duke.usask.ca
> 
> Thank you
> 
> Peta Bonham-Smith
> Department of Biology
> University of Saskatchewan
> Saskatoon
> Canada



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