Lambda DNA prep
ckwen at expert.cc.purdue.edu
Wed Jun 12 11:07:19 EST 1996
Thanks John for posting your lambda DNA prep by using DE-52. I got
several questions regarding the preparation of DE-52 in your
1. Why the MgCl2 is important in the preparation of DE-52 after the HCl wash?
I understand that HCl washing is an important step in "precycling" of
DE-52, a DEAE-cellulose, but do not know the function of MgCl2 in the
following step. As a standard "precycling" procedure of DEAE-cellulose,
the DEAE-cellulose is washed by HCl and then by NaOH, and the resulting
counter-ion will be OH-, and the salt conc will be 0.5M of NaCl. To remove
the salt which will interfere with the binding capacity of DEAE, low-salt
buffer is applied to the resin.
2. Is it a procedure modified from an article published from
Nucleic Acids Research several years ago? Anyway, your procedure is
more improved in the aspect of ion-exchange.
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