Binding DNA templates to agarose beads for invitro transcription

warwick at staff.monash.edu.au warwick at staff.monash.edu.au
Thu Dec 15 21:31:56 EST 1994


G'Day!

I want to immobilise a DNA template to agarose beads via a biotin labelled 
nucleotide.  The aim is to fill in one end of the template with the 
nucleotide-biotin, bind this to agarose-streptavidin beads and then use the 
template for in vitro transcription assays (on plant extracts).

I need help/ideas on the following:

1.  suppliers of avidin-agarose or streptavidin-agarose suitable for this 
purpose. (or magnetic?)

2.  Why is biotin -dUTP used in preference to other dNTPs?

3.  suppliers of biotin labelled nucleotides.

4.  any protocols or suggestions.

Please respond by Email.

Cheers.

Warwick

Dept of Genetics and Developmental Biology
Monash University
Clayton  VIC  3168
Australia

WARWICK at CCS1.CC.MONASH.EDU.AU




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