Fillling in/Chewing back/Blunting - will this work?

Keith Rand Keith.Rand at molsci.csiro.au
Tue Aug 11 02:31:02 EST 1998


In article <grovesa-0808982021010001 at 131.215.121.118>,
grovesa at cco.-removespam-.caltech.edu (Andy Groves) wrote:

> A simple question for you. Will this reaction work?
> 
> 
> BamHI end:      G A T C C N N N N N N .....
>                         G N N N N N N ......
> 
> 
> 
> Add Klenow and A, T, G, but *not* C
> 
> 
> 
> to give           A T C C N N N N N N .......
>                   T A G G N N N N N N ........
> 
> 
> ....i.e., filling in three bases and chewing back the lone G to give a
> blunt end.
> 

You would have to use Mung Bean nuclease (or S1 at low temperature) to
remove the 5' G after Klenow has partially filled-in.

-- 
Keith Rand,  Sydney Australia



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