sequencing from 300-500 bp

Michael Benedik bchs1b at Elroy.UH.EDU
Thu Nov 12 01:19:33 EST 1992


In article <Af0KlOu00iV005CmZv at andrew.cmu.edu>, "Morris F. Manolson" <mm6y+ at andrew.cmu.edu> writes:
>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 

You may be thinking of the Long Ranger gel reagent. It is sold by
J.T. Baker (the big chemical company). I don't know if they actually make
it or buy from someone else. You can get it from VWR and probably from other
companies also. 

I have never tried so don't take this as a recommendation...


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 Michael Benedik				INTERNET: Benedik at uh.edu
 Dept. of Biochemical & Biophysical Sciences	
 University of Houston				BITNET: Benedik at uhou
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