Taq t-tailing & Mg concentration.

pmiguel at bilbo.bio.purdue.edu pmiguel at bilbo.bio.purdue.edu
Sat Feb 12 11:56:17 EST 1994


In article <2jdruo$jag at mserv1.dl.ac.uk>, txpljfg at UABCVSR.cvsr.uab.edu writes:
...
>  Lately, we have been making our own T-vector
>from pUC19 by cutting it with smaI and tailing it for three hours with
>taq pol  at 75C in standard 1X PCR buffer (w/ 1.5mM MgCl2) containing
>2mM dTTP.  Phenol extract it, ether extract it twice (hey..so I'm
>paranoid), ppt it w/ ammonium acetate and isopropanol.  We have found
>that our vector gives slightly higher backgrounds (probably due to
>incomplete cutting).
    
  Someone mentioned to me that at 2mM dTTP, virtually all Mg ions would
be chelated.  Does Taqs tailing ability work better at low Mg ion 
concentration?

Phillip



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