Beckman sequencer problems
t.vink at med.uu.nl
Fri Nov 3 07:42:01 EST 2000
On Tue, 31 Oct 2000 11:40:53 +0000, Ivo Rieu <irieu at sci.kun.nl> wrote:
>Hello, I would like to come in contact with people that are using a
>Beckman CEQ2000 sequencer for DNAseq.analysis.
>The problem we have here is that the result is very variable. If we do
>the same sequences multiple times, one may be very nice, the other crap.
>We have the feeling that we are working on the edge of what the
>sequencer can handle, so sometimes it is just allright, while the next
>time the sample we try to sequence is just out the range the sequencer
>can handle. And we don't know wether what is out of range is the amount
>of template, the salt or something else that is not optimal. The most
>observed thing that goes wrong is a unstable, decreasing current, and a
>very late loading of the sample on the column (what is cause and what is
>- Any idea what we do wrong ?
>- Maybe someone has made some modifications to the original protocol
>that improve the sequencing ?
>Dept. Plant Cell Biology
>University of Nijmegen
>irieu at sci.kun.nl
This smells like overloading. Try diluting the sample (1:5- 1:10) or
reduce the injection time and run again.
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