Lambda restriction problem

sarven sarven at ecc.ubc.ca
Tue Sep 28 21:32:59 EST 1993


Subject: Lambda restriction problem
From: kavousi
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
the
>enzyme EcoRI but under normal condition I get very small amount of
restricted
>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?

- Sarven



More information about the Methods mailing list