Creating T-cloning vector
Dr Antony Halsall
halsalla at cf.ac.uk
Thu Feb 18 12:27:09 EST 1999
Hi
I am trying to produce a T-cloning vector for the direct ligation of a
PCR product into a reporter plasmid (pGL3-basic). I know it is possible
to create the necessary 3' T overhangs by incorporating two adjacent
restriction sites for either HphI, AspEI or XcmI, however all these
enzymes will cut pGL3-basic. So my question is does anyone know of
another enzyme that will allow the creation of 2 3' T overhangs or a
method for cutting at a site present in the pGL3-basic and filling in to
create the appropriate overhang?
Thanks in advance
Dr A. Halsall
Dept. Psychological Medicine
UWCM
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