Can a linearized Plasmid DNA ligate inside the E.coli
tfitzwater at somalogic.com
Tue Nov 12 10:19:04 EST 2002
Can a linearized Plasmid DNA ligate inside the E.coli?
My experience is yes. In 1981 I cut a ColE 1 derivative to completion with
Hind III and then lightly digested the linear plasmid with Bal 31. Most of
the ends were intact and only a small percentage were chewed back in order
to limit the extent of digestion. The DNA was ligated and then recut with
Hind III to eliminate the DNA that retained the Hind III site. This was
transformed into C600 cells and resulted in hundreds of colonies. The
problem was eliminated by dephosphorylating the recut Hind II ends before
the transformation step.
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