Antarctic Phosphatase

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This is the procedure we tried for phosphatization:

Step 1: Add 1/10 volume of 10x Antarctic Phosphatase Reaction buffer to 1 ug of DNA cut w/ any restriction endonuclease in any buffer.

Step 2: Add 1 ul (~5 units) of (A-Pase)/mix.

Step 3: Incubate for 15 min. @ 37 C for 5' extens. or blunt-ends; 60 min. for 3' extens.

Step 4: Heat inactivate for 5 min. @ 65 C (or as required to inactivate the restriction enzyme).

Step 5: Proceed w/ ligation.

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