ligation of bluntet fragments
Trygve Meum Eliassen
t.m.eliasen at bio.uio.no
Thu Apr 23 07:21:06 EST 1998
I'm having difficulties cloning a PvuII-SmaI fragment (i.e blunt in both
ends) into a vector cut with EcoRV (blunt) and AatII (3' overhang). The
sticky end has been bluntet with T4 DNA polymerase (NEB). I figure the
problem is inefficient blunting and/or ligation. Has anyone got any
suggestions as to how I might improve any of these methods? I'm mostly
following the instructions in the NEB catalogue.
Thanks in advance. thomas.moen at biotek.uio.no
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