Dear DNAseq'ers,
we are trying to get sequences from the 3'end of cDNAs in a
medium-throughput manner on our 3700. We had been using a "T(20)VN"
primer (I mean twenty T's with (ACG)(AGCT) at the
3'-end) with some success.
But all of a sudden we got the following problem: The sequence is
more or less unreadable due to a series of T signals which disappear
after about position 200 in the read. The signal intensity of these
T-peaks varies. Also, the spacing of the T-peaks is different to that
of the "real" sequence. We do not know what we might have changed
compared to the "winning team" before ....
There are pictures of the problematic reads at:
http://adis.mpiz-koeln.mpg.de/Adis/Example-Gels/read-Tstrech-T20VN.html
Any comment is wellcome!
Thanks is advance, Bernd
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_____________________________________________________________________
PD Dr. Bernd Weisshaar
MPI fuer Zuechtungsforschung (english: MPI for Plant Breeding Research)
Carl-von-Linn=E9-Weg 10
50829 Koeln
Germany
Tel. office: +49-221-5062 590 Tel. lab: +49-221-5062 311
Fax: +49-221-5062 313
mailto:weisshaa at mpiz-koeln.mpg.de
WWW-URL: http://www.mpiz-koeln.mpg.de/~weisshaa/BW-info.html
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