Effect of TE on ligations?

Andrew Doherty A.Doherty at Bris.ac.uk
Mon Dec 2 03:29:24 EST 1996


Kevin Mulcahy wrote:
> 
> Does anyone know if the presence of TE buffer in a ligation reaction can
> affect the reaction causing ligation failure? If so, how much TE (in a
> 20µl reaction) would cause inhibition, or is more appropriate to
> dissolve the DNA fragments in water in order to avoid the presence of TE
> in the ligation reaction? Lastly, if water is recommended, should I
> adjust the pH of the water to 7.5-8.0 (since our lab water is pH 6-6.5)
> so that the DNA will dissolve more easily?
> 
> Many thanks for your help,
> 
> Kevin Mulcahy.
TE shouldn't be too much of a problem, but if you're worried about it,
dissolve the DNA in just 10mM Tris, pH8. I find this works just fine.

TTFN - Andy
-- 
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Dr Andrew Doherty		email -  a.doherty at bris.ac.uk
Dept. Anatomy			Tel (0117)9287421
School of Medical Sciences	Fax (0117)9287402
University of Bristol
University Walk
Bristol UK
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