sequencing from 300-500 bp

Morris F. Manolson mm6y+ at andrew.cmu.edu
Wed Nov 11 15:26:02 EST 1992


Dear Netters,  I am sure someone must have asked this before but......

I am trying to sequence off of one reaction 300 to 500 bps using the 
standard IBI or BRL 44 cm glates.  Without having to resort to ultra-long
plates (of which I don't have), does anyone have any suggesting on how to
improve the resolution of the bands in the 300 to 500 bp region.  My reactions
are fine and I can see bands up to 600 bp from the primer, I just can't
resolve them well enough to read.  I am using 6% acrylamide wedge gels and run
them for 24 hours at 900 Volts, this gives me nice sequence on average
up to 350 bp,  on a great gel I can get 400.  I remember some company trying
to sell an alternative matrix to acrylamide, (of course I don't remember
who or where) which they claimed improved resolution, does anyone
out there know what I am talking about?  Thanks in advance, Morrie 



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