Labeling the 3′ Termini of Oligonucleotides Using Terminal Deoxynucleotidyl Transferase

P]ddATP or biotin-, digoxigenin (DIG)-, or fluorescein-ddUTP. Because none of these molecules carries a 3'-hydroxyl group, no additional molecules can be incorporated. Alternatively, the enzyme is capable of adding several (2-100) nt to 3' ends in a so-called homopolymeric "tailing" reaction. A tailing reaction is performed in the presence of a mixture of labeled and unlabeled dNTPs. The rate of addition of dNTPs, and thus the length of the tail, is a function of the ratio of 3' DNA ends to dNTP concentration and, in addition, the specific dNTP that is used.