Lambda restriction problem
sarven at ecc.ubc.ca
Tue Sep 28 21:32:59 EST 1993
Subject: Lambda restriction problem
Date: 23 Sep 93 16:23:59 +0100
In article <1993Sep23.162359.1 at mpimg-berlin-dahlem.mpg.de> ,
kavousi at mpimg-berlin-dahlem.mpg.de writes:
>I am trying to isolate a 1.4 kb fragment from a Lambda gt11 DNA, using
>enzyme EcoRI but under normal condition I get very small amount of
>fragment. It seems that the DNA is not commpletely restricted with EcoRI.
>Has anyone any idea how to improve the completeness of restriction?
Are you plating your plates using agar or agarose? It is common to lose
restriction sites if you are using agar to plate. Secondly, if you can't
cut out the insert why don't you just use the primer sites around the
insert and amplify it using PCR?
More information about the Methods