blunt ligation

[Karl Fischer]

In article <32AB60DB.7CF3 at unitel.co.kr>, "Chang,jihun"
<j6544 at unitel.co.kr> wrote:

> Dear bionetter.
>  
> I'm trying to ligate Hind III-digested genomic DNA into SmaI-digested 
> pBluescript SK vector.
> but, final white transformants I got were so few.
> I want many white colonies.
> Please, give me your help.

Hind III digestion generates a 5' overhang while Sma I generates a blunt end.

The two are incompatable unless you blunt the Hind III ends with Klenow/dNTPs.

Cheers

Karl the hepB guy

-- 
Karl Fischer
tyr-2 at bones.biochem.ualberta.ca