Non-Specificity for Primer Extension ?
leach at mbcrr.harvard.edu
Sat Mar 5 10:59:10 EST 1994
> Therefore, I am urgently seeking advice from the net about:
> 1) The possible explanation on the PE products in mock infected
> samples as well as the ways to get rid of those products (in mock).
> 2) Alternative techniques to primer extension and nuclease
> protection assays to determine the 5'ends of the transcipts.
> Thank you for your attention! I am looking forward to hearing from you.
> Jian Liu
> Dept. of Micro/Immunology
> Queen's University
> Kingston, Ontario, K7L 3N6
> E-Mail: LIU at QUCDN.QUEENSU.CA
Various solutions to this.
Try adding actinomycin D, i believe this prevents extension products acting
as primers and causing non-specific extension.
Also verify the pe products using ribonuclease protection, s1
hybridization, or RT-pcr analyses.
..... Martin Leach Email:leach at mbcrr.harvard.edu
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