cloning from agarose

Cornelius Krasel krasel at alf.biochem.mpg.de
Fri Mar 11 14:23:26 EST 1994


brett at BORCIM.WUSTL.EDU wrote:
: BOAGP at QUCDN.QUEENSU.CA writes:
: >We are doing some genomic library building and want to
: >preferentially clone fragments in the 5 to 15 kb range. We have
: >experimented with various ways to isolate this range of DNA from an
: >Agarose gel, and always seem to end up with a low yield....

: Does anybody else still do their ligations "in situ"? I have been cloning
: stuff in low melt for some time now and have never had a problem.

Me neither. I don't know if this is suitable for the construction of
libraries, though.

If it doesn't work, I just spin the LMP agarose at 100000xg, precipitate
the DNA from the supernatant and use it in my ligations. One of the two
methods always works :-)

--Cornelius.

--
/* Cornelius Krasel, Abt. Lohse, Genzentrum, D-82152 Martinsried, Germany */
/* email: krasel at alf.biochem.mpg.de                 fax: +49 89 8578 3795 */
/* "People are DNA's way of making more DNA." (Edward O. Wilson, 1975)    */



More information about the Methods mailing list