Prometa silver sequencing

Dave Ainscow d.l.ainscow at shu.ac.uk
Thu Apr 13 04:17:52 EST 1995


I am posting this on behalf of a friend of mine as she is unable
to post atm. Please send all replies to her E-mail address.

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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/andreaj at easynet.co.uk



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