PE biosystems version 3 sequencing kit

Glenis Wiebe wiebe at mpi-cbg.de
Thu Jul 19 08:16:15 EST 2001


Hi Joanne,

I was very glad to see your message.  I am having exactly the same
problem - "an offset/shift" of bases causing trouble with the base
calling.  What I see primarily are N's being called where there are no
bases; usually between about 200 and 400 bases.

It is not a drop in resolution - the peaks are still well defined.  I
think it is a problem with the mobility file, because if I analyze the
sample with the BDv2 mobility file (DT{BD Set Any-Primer}), that region
is fine.  (Unfortunately, the peak shifts still occur near the beginning
of the sequence.)

To make things even more complicated, sometimes the problem is not as
severe as others.  For the record, I always set up duplicates with the
BDv2 kit, and those results are "perfect".

Sorry, I know this is not a solution, only a conformation that you are
not alone.  I hope someone has some ideas how to fix this.

Glenis


Joanne Gilfillan wrote:

> PLEASE HELP!!!
>
> We recently tried out the Big Dye Terminator Kit Version 3 on our ABI
> 377 sequencer.  We experienced an offset/shift of the GTP in relation
> to the other bases, creating problems with the base calling. Version
> 1 of this kit gave us no problems at all. Has anyone else also found
> this, and if so, are there any ideas on how to remedy this problem?
> Does anyone know whether version3 has been tested extensively on
> gel-based systems?
>
> ---

--
Glenis Wiebe, M.Sc.
DNA Sequencing
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108
D-01307 Dresden
Germany

Tel: +49-351-210-2628
Fax: +49-351-210-2000
e-mail: wiebe at mpi-cbg.de
webpage: www.mpi-cbg.de



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