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Vector Fill in for Blunt End Cloning

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#1 Euclid



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Posted 24 March 2009 - 08:14 AM

I have a blunt ended insert I need to clone into a vector that was cut with a sticky end restriction enzyme. Using a blunt end enzyme to cut the vector is not an option. I cut the vector and filled in with klenow. Do I need to treat the linear, filled in vector with CIP? I do not think there are phosphates on the ends after fill in, because when I do this with to an insert I need to add phosphates with PNK.

Anybody have experience with this technique and able to share it?


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