MSALMINEN%FINNPHI@PUCC.PRINCETON.EDU (MIKA SALMINEN) (08/01/90)
Hello fellow netters!
Has anyone done ligations in the presence of BamH1 restriction enzyme? I have
two fragments that both have a BamH1 and a Stu1 site in ther opposite ends. As
you know Stu1 produces blunt cuts and BamH1 recessed 3':s. I want to join these
fragments at the Stu1 site without getting ligation at BamH1 (and concatamers).
In theory this should be easy if i have BamH1 in the ligation reaction, but as
i haven't tried it before, i'm a bit concerned about getting star activity by
BamH1. Is there anyone out there who knows if BamH1 cuts unspecifically in +15
C at low salt concentrations (ligation buffer) and long incubations (24-48 h),
and if so, how i could awoid this.
Thanks already, Mika Salminen
e-mail: MSALMINEN@FINNPHI (BITNET)
Adress: Finnish Public Health Institute
HIV-lab
Mannerheimintie 166
00300 Helsinki
FINLAND