<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
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