Is it OK to ligate at RT overnight?

Paul N Hengen pnh at fcsparc6.ncifcrf.gov
Wed Sep 20 17:28:05 EST 1995


 Andrei Popov <ANDREI.POPOV at bbsrc.ac.uk> writes:

| There was a paper in NAR a couple of years ago showing
| that ligations in the ultrasonic bath  go quicker
| presumably because of the frenetic movements of the DNA ends.

Some people have argued that the correct controls for the experiments published
in that paper were not done. In fact, the exact conditions cannot be duplicated
because the authors did not specify them. I wonder if anyone has done any
experiments where DNA has been added to cells and this mixture subjected to
ultrasonic frequencies. Why do most transformation protocols have a wait period
after adding DNA? Is it to let the DNA diffuse and stick to cells?  Anyone know
of studies done on speeding this up?

@article{Cooper1993,
author = "J. B. Cooper
     and D. L. Halverson
     and C. Coldren",
title = "Ultrasonic ligation for rapid high efficiency
subcloning in plamsid vectors",
journal = "Nucleic Acids Res.",
volume = "21",
pages = "1681",
comment = "ligations in an ultrasonic cleaning bath",
year = "1993"}

@article{Hengen1993Novtibs,
author = "P. N. Hengen",
title = "Methods and reagents - Bummer buffers and lightning
ligations",
journal = "Trends in Biochemical Sciences",
volume = "18",
number = "11",
pages = "446-448",
month = "nov",
year = "1993"}

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