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
-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.
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
I'll appreciate any comments and opinions
thank you
-medchemgirl-