I am using a antarctic phosphatase and a line in the protocol says, '' incubate for 15 minutes at 37 deg C for 5' extensions or blunt ends, 60 minutes for 3' extensions. ''
Does it mean, for 15 minutes it will remove 5' phophate groups and in a longer incubation of 60 minutes it will remove 3' groups (which i think is hydroxyl or is it phosphate too in some conditions?). i jjust want to prevent vector religation before i put the insert. Pls help understand this line in the protocol.
You all have always been very helpful, esp. veteran moderator, and am very thankful, thanks in advance again.