48cm plates on the ABI 377

Rick Nicholson R.Nicholson at arnie.cfi.unsw.EDU.AU
Fri Aug 9 02:49:41 EST 1996


Derek Potter wrote > We have been trying out the 48cm plates on our 
377 and our results are very
> similar to those obtained with the 36 cm well to read system @ 2X conditions.
> I was wondering if anybody had any success by playing around with the run
> conditions or Long Ranger etc?
> 
This question sounds very similar to my question posted here in late 
July.  From the responses I got it is clear that a lot of us have the 
same problem.  Several good helpful suggestions were also given 
though.  I will forward the responses people sent on to you directly.

In summary the way to get longer reads of good sequence with the 48 
WTR is to drop the run temperature to 48C and change from standard 
19:1 acryl:bis to a 29:1 ratio.  The people at FMC emailed me to say 
that LongRanger does not help in 48 WTR gels but they are working on a 
new formulation which will.  I have obtained samples of different 
acryalmide mixes such as 29:1, 37.5:1 and PagePlus which I am trying 
to see which works best for us.  So far I can say that dropping the 
temperature and changing to 29:1 acryl:bis has helped enormously.  

-- 
Richard C. Nicholson, BSc PhD        R.Nicholson at cfi.unsw.edu.au
Senior Research Fellow
Centre for Immunology
St. Vincent's Hospital
Sydney, Australia 2010




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