PCRing 5' UTR using a cDNA plasmid?

Shahram Mori smori at nmsu.edu
Wed Nov 16 02:11:09 EST 1994


Mari L. Shinohara (Mari.L.Shinohara at dartmouth.edu) wrote:
: Hi folks,

: I need to amplify 5' untranscribed region from genomic (total) DNA. 
: However, I now only have a cDNA plasmid of the gene of interest.  I
: heard that there are some ways to do it by using anchor(s) (not really
: RACE in this case).  Could you send me any protocols and/ or ideas? 
: Your help will be appreciated!

: Mari

You have the cDNA. Why don't you do a primer extension reaction? Other
wise you would have to digest the genomic DNA, ligate a primer of your own
choice to it and do a PCR using the anchor primer and down stream primer (
using your cDNA sequence of course.
Cheers,
		 	 
--
Shahram Mori					   _/\_
Program in Molecular Biology			  _\  /_
Dept. of chemistry and Biochemistry Box 3C	  \_  _/
NMSU  Las Cruces NM				    ||
88003





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