Talk:Phosphatase treatment of linearized vector

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I used this protocol when cloning a piece of DNA with SpeI and XbaI ends into a vector with SpeI and XbaI ends. Since SpeI and XbaI form compatible ends, the self-ligated vector, produced via an intramolecular ligation event, is highly favored over the bimolecular ligation event. However, when the vector was treated with this protocol, no transformants were obtained on the vector only control plate suggesting that this protocol is very effective at reducing background. In the absence of phosphatase treatment, many transformants were obtained so this step is necessary for obtaining transformants with the proper insertion. --Reshma 14:31, 7 Jun 2005 (EDT)