CIP required for double digestion of plasmid?
Posted 05 March 2009 - 06:13 AM
Posted 05 March 2009 - 07:13 AM
If i am doing a double digestion, is the calf alkaline phosphatase required?
Technically, if the vector is double digested, CIP is not required. It can be skipped and some people do.
Personally I always dephosphorylate my vector. It is an extra safe guard should the digest of the vector fail to go to completion.
However one must be careful when using CIP as over dephosphorylation will damage the end of the vector, preventing ligation.
Posted 05 March 2009 - 07:59 AM
Posted 05 March 2009 - 08:17 AM