deletion by PCR

Andrew Schrader andrews at
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?


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

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