<None>

Martin Kennedy mkennedy at chmeds.ac.nz
Thu Mar 3 21:12:36 EST 1994


In article <1994Mar1.052847.8742 at alw.nih.gov>, bernard at elsie.nci.nih.gov (Bernard Murray) writes:
> In article <smrodems-280294222013 at f180-045.net.wisc.edu>, smrodems at students.wisc.edu (Steve Rodems) writes:
>> In article <vernon.16.0009A61A at micro.uct.ac.za>, vernon at micro.uct.ac.za
>> (Vernon Coyne) wrote:
>> 
>> I have heard that borate can cause DNA damage so that some people stay away
>> from borate containing buffers when the want to use the DNA for further
>> cloning. 

> The one time you can't
> use TBE is when you intend to gel purify a fragment for ligation as the
> borate inhibits the ligase (I don't know if this means *all* ligases).
> Then I use TAE and run the gel overnight at low voltage.
> Bernard Murray, Ph.D.

This is news to me, as for over 10 years now I've been running solely TBE based
gels for everything, including fragment isolation, and I've made numerous 
recombinant constructs.  Presumably borate is only a problem if it comes 
through the precipitation step, and I'd have thought the 70% ethanol wash would
take care of this.  Any thoughts on this?
-- 
Cheers,

Martin

NNNN   NN  Martin A Kennedy (E-mail = mkennedy at chmeds.ac.nz)  ZZZZZZZ  
NN NN  NN       Cytogenetic and Molecular Oncology Unit          ZZZ
NN  NN NN           Christchurch School of Medicine            ZZZ
NN   NNNN              Christchurch, New Zealand              ZZZZZZZ
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