Discussion of DNA Sequence

scole at welchlink.welch.jhu.edu scole at welchlink.welch.jhu.edu
Fri Jul 11 03:36:27 EST 1997


One suggestion -- If you did this TA cloning from a commercial TA cloning
kit kit purchased relatively recently (last year or so, I think?) you
should be aware that among the changes Invotrogen made between the
original TA vector and vector pCR2.1 was the removal of the SP6 site
altogether.  If you're cloning into pCR2.1 you should use M13r to sequence
from that end.

Susan
scole @welchlink.welch.jhu.edu

In article <33C55CA1.3E9E at ns.east.cn.net>, ddsyr at ns.east.cn.net wrote:

> Dear Netters:
> 
> I have met many problems in DNA sequence recently, so I like to initiate
> a discuss on this topics. I cloned a PCR product and insert into a
> T-vector. The enzyme check of polylinker is OK. The sequence from one
> end (T7) by DNA automator is OK. But the sequence from another end
> (SP6)did not get  any signal. The sequence was conducted by a commercial
> company since our lab has no such equipment. Could any one give your
> idea what is wrong about it? I also like to know the general
> troublesomes in DNA automation sequence and how to avoid them? Any
> seggestion and discussion is greatly welcome.
> 
> Dr Q. H. Ma
> Institute of Botany, Academia Sinica
> Beijing 100093, China



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