DNA Seq; Nasty template

Martin Pentony (mpento@cmgm.stanford.edu)
Fri, 5 Jun 1998 16:59:24 -0700

Hi, a customer gave me two templates based on a "pOX"? vector. We
sequenced with T3 and T7. The T3 worked fine but the T7 had a
significant reduction in signal strength about 25 bases into sequence
after a poly G. The insert is about 200 bases from the priming site.
Below is the first 50 bases of sequence. It is not reliable after the
poly G. Any details on this vector and is this drop in signal due to
the poly G or some other secondary structure and what are the possible
strategies to sequence through this.

<bigger><bigger> 1 CGANTGGNGC TCCACCGCGG TGGGCGGCCG CGGNGGGGGG
GGGGGGGGGT

51 TTTTTTTTTT TNTTTNCNTT TTNTTTNGAA TGNGAACAAA CTNTGTTTTT

</bigger></bigger>thanks for the help

Martin Pentony

DNA Sequencing section

P.A.N. Facility

Stanford University

http://cmgm.Stanford.EDU/pan/