HELP : Blunt to Sticky-end ?

Shane Albright insomnia at mendel.berkeley.edu
Fri Jan 27 22:19:06 EST 1995


In article <1995Jan19.145157.7473 at nki.nl>, gbrink at nki.nl wrote:

> Dear All,
> 
> Who can tell us how to link one single T to a blunt-end DNA fragment, to
create
> a sticky T-overhang ?
> 
> Guido Brink &
> Arno Floore
> Netherlands Cancer Institute, Amsterdam

Incubate the DNA with Taq polymerase (1 U/ ug plasmid/ 20 ul volume) and 2
mM dTTP in 1X Taq buffer for 2h @ 70°C.  Then phenol extract and EtOH ppt.

See also Marchuk et al, Nucleic Acids Res 19(5), p. 1154.

Shane Albright
MCB Dept., UC Berkeley
Berkeley, CA



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