Non-Specificity for Primer Extension ?

Martin Leach 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
>                             Canada
>                             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.
-- 

.....          Martin Leach                Email:leach at mbcrr.harvard.edu 
   _|____      Dept. of Pharmacology       Phone: (617) 638-5323        
   / o  /      Boston Univ. School of Med. Fax:   (617) 638-4329         
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