PCRing 5' UTR using a cDNA plasmid?

[Mari L. Shinohara]

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

[Shahram Mori]

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

: 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