Promega Silver Sequencing

Andrea Johanson andrea.johanson at nri.org
Mon Aug 21 13:36:42 EST 1995


Does anyone have experience with Promega's Silver Sequencing system...?

We have tried unsuccessfully for far too long to get this system to work in our laboratory.  I'm still unsure whether it is a
staining or a gel-running problem, and don't have enough experience of either to be able to judge.  So far we have only
used the control template DNA supplied with the kit, and we have used several kits with different batch numbers.
We have played with almost (if not every) parameter imaginable, but are unable to get any sequence information.

Instead of the nice clear bands (I have seen a gel that worked so I believe it does work!), we get smears of DNA in the
loaded lanes.  Even the markers smear, which maybe suggests it is a gel problem, but we have tested all our buffers etc
extensively.  Towards the bottom of the gel we can often see faint, diffuse bands (with a little imagination - so important in
science..), but nothing to get excited about.

Promega have been very helpful, but are also baffled as to what is going on.  Any ideas, however bizarre would be much
appreciated!

Thanks.

Andrea Johanson
Natural Resources Institute
andrea.johanson at nri.org





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