T overhang

Dom Spinella dspinella at chugaibio.com
Thu Jun 26 12:39:33 EST 1997


> How do I generate a 5' overhang on a SMA1 cut site so that I can clone a 
> PCR products with 3' A overhang? Thanks for the info. I do know this is 
> old stuff. It's just my memory is not very good. Bye for now.
> Chong

Chong:
There are several methods to roll your own T vectors, and you need only
go back through this month's and last month's postings to learn about
them (if you just search my name, you'll find my response and several
others to the same question).  However, if you're set on using a SmaI
site,  wouldn't it be easier to just blunt your PCR product with Klenow
or T4 Pol (or just generate the PCR product in the first place with Pfu
or some other thermostable polymerase with 3' -> 5' exo activity) rather
than go through the whole exercise of generating a T-vector?  Ayyway,
just a thought.  Good luck.  -- D.G. Spinella



More information about the Methods mailing list