5' overhang repair

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Sat Jul 15 13:36:55 EST 2000


Hi Ranga,
I´m not sure if I understand your question right, but if your
"mismatches" are a 5´ overhang of the insert (that don´t match the
vector), you simply don´t have compatible ends. These won´t clone at
all. Sounds like a bad strategy.

Frank


Ranga Uday Kumar wrote:
 
> Dear netters,
>     A cloning strategy will result in a recombinant plasmid with a two
> base pair-mismatch at both ends of the insert. The two base
> pair-mismatch is a 5' overhang at both the ends. Will the mismatched
> bases be repaired by the standard E.coli hosts like DH5 alpha? Or is it
> essential to repair them in vitro before transfection? Which is the best
> repair strategy if it is required? Your suggestions are highly
> appreciated.
> 
> --
> ________________________________________________________
> Ranga Uday Kumar Ph.D.
> Faculty Fellow
> Molecular Biology and Genetics Unit
> Jawaharlal Nehru Center for Advanced Scientific Research
> Jakkur Post, Bangalore 560 064
> India
> Tel: +91 (80) 846 2750 Ext. 241
> Fax: +91 (80) 846 2766
> ________________________________________________________
> 
> ---






More information about the Methods mailing list