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phosphorylated oligos and vector - (Sep/08/2005 )

my vector is cut with 2 different RE and is not CIP treated. the ds oligo insert is also phosphorylated. does having phosphorylated ands on both vector and insert affect ligation efficiency?


As long as you do not de-phosphorylate (CIP) your vector, there is no need to phosphorylate your oligo, although i quess it want harm. But ofcourse, you will have to make your oligo double-stranded.


well assuming oligos are phosphorylated in 5' and the vector too, it's the best conditions for ligation. if you encounter too much self ligation of the vector you will need to CIP treat your plasmid.