IS IT OK TO LIGATE AT RT OVERNIGHT?

Volker Knoop knoop_v at mpimg-berlin-dahlem.mpg.de
Tue Sep 19 14:08:14 EST 1995


In article <43lfht$ilk at nntp.ucs.ubc.ca>, bina at unixg.ubc.ca (rew) wrote:

>I DO NOT HAVE MUCH EXPERIENCE DOING LIGATIONS AND I HAVE TWO QUESTIONS:
>
>1. I DO NOT HAVE ACCESS TO A 15C INCUBATOR AND WAS WONDERING IF ANYONE
CAN TELL 
>ME IF THER IS A PROBLEM WITH DOING LIGATIONS AT RT OVERNIGHT?
>
>2. IS THERE A PROBLEM USING HIGH CONCENTRATIONS OF LIGASE (1-3 UNITS)IN MY 
>LIGATION REACTIONS?


1. No, RT overnight is perfectly fine and even the temp of choice for
blunt ligations. Decreasing temperature makes sense for sticky ends.
2. So, what is the concentration you use? 1-3 Units in 10 mikroliters is
generally fine.

-- 
Volker Knoop
IGF Berlin Dahlem
14195 Berlin
phone +49-30-830007-48 Fax -36
e-mail: KNOOP_V at MPIMG-BERLIN-DAHLEM.MPG.DE



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