Protocol Online logo
Top : Forum Archives: : Molecular Biology

Is it ok to store a digested vector for ligation? - (Jun/24/2005 )

Would you recommend to digest the vector right before ligation? or it could be store at -20 ready to ligate, to be used when needed?

I'll appreciate any comments and opinions
thank you
unsure.gif

-medchemgirl-

If I am going to do a whole bunch of ligations in the same vector, using the same cloning scheme (same restriction digests), I'll cut up a bunch of vector, purify it and store it at -20 and thaw and reuse as needed.

Did13 Not/Kpn ligations into the same vector this week with this thaw and use method. Worked like a charm.

-pBluescript-

QUOTE (pBluescript @ Jun 24 2005, 07:05 PM)
If I am going to do a whole bunch of ligations in the same vector, using the same cloning scheme (same restriction digests), I'll cut up a bunch of vector, purify it and store it at -20 and thaw and reuse as needed.

Did13 Not/Kpn ligations into the same vector this week with this thaw and use method.  Worked like a charm.


I also used to do the same way. Made a large amount of stock and used for several times later.

-dtle-

hi
i don't see any contradiction in make a store of digested plasmid. As pBluesciprt and dtle suggested. I would recommend to do large stock but in several aliquots to minimize freeze thaw repetitions.
fred

-fred_33-

Thank you !!

QUOTE (medchemgirl @ Jun 24 2005, 04:11 PM)
Would you recommend to digest the vector right before ligation? or it could be store at -20 ready to ligate, to be used when needed?

I'll appreciate any comments and opinions
thank you
unsure.gif

-medchemgirl-