A 1 What is the point of using calf intesting phosphatase 2

A. 1. What is the point of using calf intesting phosphatase?

2. What is the ideal condition for CIP enzymatic activity?

3. Does CIP work in conventional restriction enzyme buffers?

Solution

1 ANS- Calf intestinal alkaline phosphatase or CIP Catalyze the removel of 5\' and 3\' phosphate group from nucleic acid i.e. DNA and RNA .so that it can be replaced with radiolabelled phosphate .it also remove phosphate group from ribo and deoxyribonucleoside tri phosphateIn cleaved DNA CIP is very useful for the prevention in self ligation of cleaved DNA vectors because the lack of 5\' phsophate group can not ligate.it also helps to prepare template in DNA sequencing in PCR reaction.

2 ans-CIP are zinc containing enzyme possesing one zinc atom and inhibited by EDTA which acts as a chekating compound.

3 ans-CIP acts on NEbuffers 2,3,or 4 which is optimal but should not used in NE buffer 1 for Eco R1