4 way ligations?

[Ned Mantei]

In article <DtDCtJ.2pI at news.hawaii.edu>, angel at ahi.pbrc.hawaii.edu (Angel
Yanagihara) wrote:

> Hello...
> 
> I'm attempting a 4 way ligation and was wondering if anyone out
> there has had any luck with this kind of thing.

Even a 5-way ligation is possible. Try about 10 femtomol of each fragment
in a 10 ul ligation reaction, overnight with about 1 Weiss unit ligase. I
don't think there is anything gained by ligating some segments separately.
You should be getting 10's to 100's of clones with anywhere up to 95%
being correct.

-- 
Ned Mantei
Dept. of Neurobiology, Swiss Federal Institute of Technology
CH-8093 Zurich, Switzerland
mantei at neuro.biol.ethz.ch   Fax: +41-1-633-1046