Rapid Ligation Kit

David L. Haviland, PhD dhavilan at imm2.imm.uth.tmc.edu
Wed Jan 5 20:12:59 EST 2000


On 6 Jan 2000, David Mckinnon wrote:

> We have been using Boehringer Mannheim's rapid ligation kit for some time now
> and find it to work very well. It cuts a day from most cloning procedures and
> gives a very high and reproducible yield of recombinants. Does anyone have a
> home-brew procedure that works as well as the kit? It is unlikely that
> Boehringer has stumbled onto something very novel but our efforts to replicate
> their kit have not worked as well as the proprietary reagents.
> 
> Thanks for any suggestions

In what regards?  What part of a standard ligation is truncated with this
kit?  

With sticky end ligations, I've been able, on more than one occasion, to
prep vector and insert, add standard ligation buffer and ligase, incubate
on the bench (room temp) for 30 mins to an hour, electroporate, and be
looking at colonies the next morning. 

So again, I ask, what part of the "standard" ligation protocol is reduced?

David
==========================
David L. Haviland, PhD
Assistant Professor, Immunology
University of Texas, HSC - Houston
Institute of Molecular Medicine
2121 W. Holcombe Blvd.
Houston, TX  77030
http://www.uth.tmc.edu/~dhavilan
713.500.2413-Voice
713.500.2424-FAX
==========================



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