5' RACE Specificity Problems
smori at nmsu.edu
Wed Nov 16 02:16:16 EST 1994
Sarah Childs (childs at oci.utoronto.ca) wrote:
: I'm having trouble with specificity in 5'RACE.
: I'm getting non-specific bands, even though my gene-specific
: primer is an 18 mer with a Tm of 58C, I'm doing the PCR with a hot start and
: annealing at 56C. At the same time I am not getting the product I want.
: Has anyone had similar problems and overcome them? Does anyone have a
: very good RACE protocol that might help me?
: Sarah Childs
: Ontario Cancer Institute
: email: childs at oci.utoronto.ca
The problem is probably at your anchor ligation Sarah. I used Clontech's
5'RACE but I never got anything. For very few people it worked.I would
make sure that you are getting your anchor ligated to the 5' ends.
Shahram Mori _/\_
Program in Molecular Biology _\ /_ Saskatoon/SK/CANADA
Dept. of chemistry and Biochemistry Box 3C \_ _/
NMSU Las Cruces NM ||
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