Isolating areas flanking a gene

Brad Nicholson Brad_Nicholson at hlthsci.med.utah.edu
Mon Oct 30 17:34:30 EST 1995


In article <4738s1$qiq at er6.rutgers.edu>, driscoll at eden.rutgers.edu (William
Driscoll) wrote:

> I would like to isolate the areas flanking a particular cloned gene from 
plant 
 >cells. 
>strand isolation idea sniped< 
 >Also any other alternate ideas for isolating the areas 5' of a gene would
be  >interesting to hear. 
 >
 >Thanks in advance,
 >Bill
 >driscoll at eden.rutgers.edu

Hi Bill,
Have you considered inverse PCR (IPCR)?  I have tried it to isolate DNA 
flanking a Tn10 insertion in bacteria.  WHile plant genomes are much more 
complex, it may work.  You can do a literature search to see if anyone has 
used IPCR in plants or you can look at some stuff which has worked in 
bacteria.  There is a reference for inverse PCR off of Tn10 in Infect. 
Immun. (1994) 62:1623-1630.  This is what I used as a starting point and it
worked, first time. :)  Email me if you have questions, and I'll try to 
help you as much as I can.

Good Luck,
Brad
===========================================================================
Brad Nicholson                      |"If it worked the first time, 
Department of Pathology             |	it wouldn't be research."
University of Utah                  |	 ...Brad Nicholson
Salt Lake City, UT 84132            |  My opinions are solely my own.
Brad_Nicholson at hlthsci.med.utah.edu |
or: (801)-581-4365                  |  iligitimi non corborundrum
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