Combinations of SalI and HindIII - Conditions for double digestion (Aug/27/2008 )

Hi Members,

Have anyone of u ever used the combination of SalI and HindIII or Subcloning. If so did u perform double digestion or Sequential.
Could anyone who have succesfully used the above enzyme combination please send me the conditions for setting restriction digestion......

Thanks in advance

Clonetech

-clonetech-

QUOTE (clonetech @ Aug 28 2008, 11:47 AM)

Have you looked at the NEB website? There's a section on double digestions.
http://www.neb.com/nebecomm/DoubleDigestCalculator.asp?

-swanny-

In our lab, we usually use Promega Multi-Core Buffer, SacI and Hind III and this system works well.

-megachunk-

Are you trying to digest a PCR product? If so, Sal I is no good, because it doesn't like PCR products, according to NEB...

-swanny-

QUOTE (swanny @ Sep 5 2008, 05:47 AM)

Are you trying to digest a PCR product? If so, Sal I is no good, because it doesn't like PCR products, according to NEB...

I agree. I would even go so far as to say that SalI doesn't like anybody. It is finicky enzyme, not the best thing to rely on in a ligation strategy.

-perneseblue-