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Affinity purification of DNA-binding proteins

Stephane Vuilleumier svuilleu at micro.biol.ethz.ch
Wed Jul 13 04:11:37 EST 1994


Hi all,

I was wondering if anybody has a protocol for agarose-avidine
affinity purification of DNA binding proteins with biotin-
tagged DNA target sequences involving DNA fragments of 350 bp?
(We know of one publication where a 240 bp fragment was used:
(Chodosh et al. (1986) Molecular Cellular Biology 6, 4723-4733). 
Are there any special unpublished tricks in these experiments?
The size of agarose beads also seems important:
4-6% beaded agarose is usual. 
Can lower percentage agarose preparations be used
for our purpose and are any of these commercially available?


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Stephane Vuilleumier, Ph.D.
Mikrobiologisches Institut
ETH-Zurich
ETH-Zentrum/LFV                       Phone:  (+41) 1 632 33 55
8092 Zurich                           Fax:    (+41) 1 262 06 47
SWITZERLAND                           e-mail: svuilleu at micro.biol.ethz.ch
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