DGGE - help please

Jason Affourtit affouj at rpi.edu
Mon Jan 8 10:41:11 EST 1996


hi!  i have been experimenting with DGGE for quite some time now (!)
and was wondering if anyone out there would be willing to share some
of their experiences with me.  I've been using a standard Biorad Protean
II (can't afford the D-Gene :( ) with a peristaltic pump to circulate
buffer (1X TAE) and an immersion heater at 60C.  Anyone using a similar
setup or have suggestions?  one thing i've noted is in runs >3 hrs the
bands all get smeary (very) and are basically useless.  my feelings are
that this is most likely buffer depletion (using 4L 1X TAE), but anyone
else offer some suggestions?  i've seen papers with extensive run times
but their data seem very nice.  

any help at all with this would be extremely appreciated, responses by
email would be great, i can clear up any ambguities then also.

thanks very much in advance!

-jason affourtit

BTW - i am using PCR product of 359bp + 57mer (inc. GC clamp) primer and
10mer primer.

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