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Terminal Deoxynucleotidyl Transferase (TdT) is an enzyme that catalyzes the repetitive addition of mononucleotides from dNTPs to the terminal 3′-OH of a DNA initiator, accompanied by the release of inorganic phosphate. The enzyme thus provides a unique method for the labeling of the 3′ termini of DNA.

This technical bulletin provides two different protocols for the incorporation of 32 P. The first illustrates the addition of an [alpha-32 P]dNTP “tail” to the 3′ termini of single-stranded DNA fragments. In addition, incorporation can be limited to a single nucleotide by using [alpha-32 P]cordycepin-5′- triphosphate. This analog lacks a free 3′ hydroxyl group, preventing incorporation of additional nucleotides.

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