Fri, 03 Sep 2010 07:41:15 +0000 bbs@protocol-online.org (BioForum) IP.Blog 60 http://www.protocol-online.org/forums/blog/10/entry-17-e-coli-ligase-is-not-the-way/
It has been a case of "I did warn you."

I have attempted to improve the efficiency of my multiway ligation by using a different ligase, a higher fidelity ligase. The only other ligase sold commercially is E.coli ligase. It is stated that E. coli...]]> Thu, 02 Sep 2010 03:26:00 +0000 http://www.protocol-online.org/forums/blog/10/entry-17-e-coli-ligase-is-not-the-way/ http://www.protocol-online.org/forums/blog/10/entry-15-ligation-5-way-and-beyond/
I have decided to find the limits of multi-way ligation. Thus far, I have determined that the techniques I am using gives me a reproducible limit of 6-ways. 7-way ligation occasionally works. And all attempts at 8-way ligation have ended in failure. My current screen size is 96 colonies..

At this point there are two obvious ways I...]]> Sat, 28 Aug 2010 01:20:00 +0000 http://www.protocol-online.org/forums/blog/10/entry-15-ligation-5-way-and-beyond/