Help with DNA electrophoresis

Martin Leach leach at mbcrr.harvard.edu
Fri Oct 1 05:59:58 EST 1993


In article <1993Oct1.091403.11081 at crc.ac.uk>, mrhodes at crc.ac.uk (Dr. M.
Rhodes) wrote:

> Help !
> 
> I am tryimg to run nondenaturing polyacrylamide gels to visualize DNA in the range 100-400 bp
> WE are using a large sequencing rig with 1 or 2mm spacers . But it aint working
> 
> Does anyone have any conditions they have used wich might help
> 
> Volts/cm
> Current (dont want it to get to hot)
> Buffers
> Acrylamide concentration (at present doing 6%)


We do a lot of sequencing in our lab, and have some experience with
nondenaturing polyacrylamide gels.


How about using 0.4mm spacers, 3-4% acrylamide gel(containting 0.5xTBE),
use 1.0xTBE running buffer. Should be approx 1500V, approx 20-30W. Run gel
for about 3-4 hours I think.

Good Luck

Martin Leach 

(ex S.0. for Tim Crow in Psychiatry in CRC/MRC - send my regards to tim)



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