deletion by PCR

Andrew Schrader andrews at pharm.usyd.edu.au
Wed Jun 10 19:09:08 EST 1998


I have designed an experiment to create a 70 bp truncation in a cDNA
species (eventually to be used in QC-PCR). Two fragments have been
generated (either side of the region to be deleted) with the 3'-region
of the first being the same as the 5'-region of the second - a 10 bp
match using a linker on the 5'-primer of the second fragment. After
denaturation, I can not get the two fragments to anneal in order to
generate a long fragment.

Is there a paper/protocol that outlines this method and is my linker too
short to allow annealing?

Thanks

Andrew
--
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Dr Andrew P. Schrader
Postdoctoral Fellow
(Respiratory Research Group)
Department of Pharmacy (A15)
The University of Sydney NSW 2006

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