What gel & conditions to use ?

Jan De Riek j.deriek at clo.fgov.be
Fri Feb 18 03:56:02 EST 2000


Dear newsgroupers,

I'am not so familiar with this newsgroup from the bio.net family, but
with my nice exprience with the
automated sequencing newsgroup convinced me to try this newsgroup.

My problem : clearly separation of PCR-fragments (300 -3000 bp) on a
denaturating gel.

We want to clearly separate radioactive labelled PCR-fragments in a
range 300- 3000 bp.
Now we use 7.5 M urea, 6% PAA, 1X TBE on 48 cm SequiGen Sequencing Cell
from BIORAD.
The run conditions are 120 W, 1800 V, 67 mA for 2.5 h hours as used for
AFLP.
Here we see nice 100 -1000 bp fragments separated.
Does anyone know some modifications tricks to separated 300 -3000 bp
fragments ?

Thanks in advance,

Jochen Dendauw

-------------------------------
Dept. Plant Genetics and Breeding (DvP -CLO)
Applied Plant Biotechnology
Caritasstraat 21
9090 MELLE
BELGIUM

TEL         +32 9 272 28 81
FAX        +32 9 272 29 01
E-mail      j.deriek at clo.fgov.be
WWW    http://www.clo.fgov.be/dvp.htm


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