CIP Treatment

CIP Treatment Protocol
CIP treatment is done to phosphatase the vector used for plasmid ligations. This is done to reduce the self ligation of a vector digested with enzyme(s) creating compatible sticky ends and hence enhancing the Signal/Noise ratio of transformations.

1.  Use 10 units of CIP per 1µg of DNA (over digesting by factor of  X)

2.	Calculate volumes

DNA µg = DNA volume * concentration

Enzyme volume = Enzyme unit/µl* # units = X [µl]

Buffer is dilution factor x dilution of the total volume.

[i.e. for 10X over digest - buffer is 10%, 3x - 30% of total volume]

3.	Order of filling

•	DNA

•	Water

•	Buffer

•	CIP

4.	Incubate for 3 hours at the specified temperature for the enzyme (37C).

5.	Keep the buffer on ice and the CIP in the benchtop coolers when on the bench.