clonetech cDNA construction kit

rabc010 at S1.CXWMS.AC.UK rabc010 at S1.CXWMS.AC.UK
Tue Sep 9 05:41:51 EST 1997


I had problem too with Clontech cDNA library construction kit
(SMART PCR cDNA Library Construction Kit, PT3000-1). I checked
cDNA after PCR aplification and that was ok but I wanted to cut
my cDNA with frequent cutter enzyme to get short fregments for
some pruposes and clone them but without success. I could not
get asingle colony. All my contorls were fine including digestion
of the plasmid. Myself too I don't beleive in their claim.
For this kit I am not sure, upto my knowledge I don't know or 
read any article about terminal transfersae activity of reverse
transcriptase as clontech claims. I was also impressed by
this kit specially "Virtual Northerns" but I am not sure of its
validity. Have you tried "Virtual Northerns"? Now I am using terminal
transfersae to tail first strand cDNA ( A tailing) and long range PCRing
with single oligodT (XhoI-T15 VN) anchored primer. this is
working very well. you might try it. If you need protocol let me
know.

narendra
nkaushik at hgmp.mrc.ac.uk



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