Any experience in improving blunt end ligation?

Nick Theodorakis nicholas_theodorakis at urmc.rochester.edu
Fri Jun 29 09:40:44 EST 2001


In <20010629125618.4754.qmail at ww02.jatek.com>, fly wrote:
>
>I want to clone a insert with a blunt and coadhesive end into PGEX-2T. I
>uesd the protocol in Promega T4 ligase insert without addition of PEG, 
and,
>however, I did not get any desirable transforminants. After I  excluded
>other causes, i plan to use PEG8000 to improve my ligation. Could you 
tell
>me your experience and recommend a quality product?

I've been using PEG in the form of the "5X ligation buffer" from BRL oops I 
mean Life Technologies oops I mean Invitrogen. If my background gets a 
little high, sometimes I do good old fashined colony hybridization to find 
the ones with inserts.

Nick

-- 
Nick Theodorakis
nicholas_theodorakis at urmc.rochester.edu

 




More information about the Methods mailing list