When is alkaline phosphatase treatment required in vector?? Is it required every time or only when blunt end cutter enzyme is used?

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Chantal Godin

In your case, i will not use it. Only when only one enzyme is use or when the plasmid is cut with two compatible enzymes (like Xho1/Sal1; BamH1/BglII; EcoR1/Mfe1...).

Amit Kesharwani

Alkaline phosphatase removes 5' phosphate groups from vector so that prevents self-ligation of the vector and facilitates the ligation of other DNA fragments into the vector.

In your case, you have two restriction enzymes therefore there is no need to do phosphatase treatment.