Biotin/streptavidin hybrid-DNA purification

Eric Lader lader at ambion.com
Fri Oct 4 21:58:49 EST 1996


In article <534dfb$rsq at babyblue.cs.yale.edu> gary vanzin,
gfv94001 at uconnvm.uconn.edu writes:
>>I'm looking for a reference/protocol to purify hybrid DNA using a 
>biotin/streptavidin system.  I'm screening 6900 arabidopsis plants, 
>each containing 1-4 insertional mutations (T-DNA).  I would like to 
>enrich my DNA preps for the T-DNA, so I can combine 100,500, or maybe 
>even all 6900 DNA preps during the subsequent PCR screen.  I believe I 
>can do this using biotinylated T-DNA as a probe, then doing 
>hybrid-select, and purification over a streptavidin column.  Any Ideas? 
> Anybody know how well streptavidin coated dynalbeads work?

I used the promega streptavidin beads for selection of biotin probe/cDNA
clones.
As long as there is a PCR step, the yield is not really a big problem.
You can label RNA or DNA with biotin by enzymatic incorporation,
photobiotin, psorelen biotin, or nick translation or random priming with
biotin nuc's.

               **my opinions are just that**
><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><
        Eric Lader Ph.D.	Senior R&D Scientist,  
                          Ambion Inc.
Unique tools for Molecular Biology Research

Phone (800) 888-8804       Fax (512) 445-7139
				  (512) 445-6979       lader at ambion.com
><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><><



More information about the Methods mailing list