Any experience in improving blunt end ligation?

Nick Theodorakis nicholas_theodorakis at
Fri Jun 29 09:40:44 EST 2001

In <20010629125618.4754.qmail at>, 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, 
>however, I did not get any desirable transforminants. After I  excluded
>other causes, i plan to use PEG8000 to improve my ligation. Could you 
>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 Theodorakis
nicholas_theodorakis at


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