PCRing 5' UTR using a cDNA plasmid?
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!
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.
Shahram Mori _/\_
Program in Molecular Biology _\ /_
Dept. of chemistry and Biochemistry Box 3C \_ _/
NMSU Las Cruces NM ||
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