Cloning and Mapping
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  • DNA Phosphorylation

    Single- or double-stranded DNA with a 5’-hydroxyl terminus has to be phosphorylated prior to ligation as 5’ phosphate groups of a DNA strand is required for ligation. A number of polynucleotide kinases, including T4 PNK (NEB #M0201) and T4 PNK (3’ phosphatase minus) (NEB #M0236) can be used to transfer the γ phosphate of ATP to a 5’ terminus of DNA.

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