Creating T-cloning vector

Dr Antony Halsall halsalla at
Thu Feb 18 12:27:09 EST 1999


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

More information about the Methods mailing list