Terminal Deoxynucleotidyl Transferase, Recombinant

Terminal Deoxynucleotidyl Transferase, Recombinant, catalyzes the repetitive addition of mononucleotides to the terminal 3´-OH of a DNA initiator accompanied by the release of inorganic phosphate. Single-stranded DNA is preferred as an initiator. Polymerization is not template-dependent. The addition of 1mM Co²⁺ (as CoCl₂) in the reaction buffer allows the tailing of 3´ ends with varying degrees of efficiency. TdT is provided with 5X Reaction Buffer: 500mM cacodylate buffer (pH 6.8 at 25°C), 5mM CoCl₂, 0.5mM DTT.

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Applications

• Tailing reactions to add complementary homopolymer tails to DNA vectors and cDNA.
• 3´ end-labeling.
• TUNEL assays.

Storage Buffer: 50mM potassium phosphate (pH 6.4), 100mM NaCl, 1mM β-mercaptoethanol, 0.1% Tween® 20 and 50% glycerol.

Source: Recombinant E. coli strain.

QC Tests: Activity, endonuclease, DNase, RNase, 3´ end-labeling, apoptotic DNA end-labeling.

Unit Definition: One unit of activity catalyzes the transfer of 0.5 picomoles of ddATP to oligo(dT)₁₆ per minute at 37°C in 1X Terminal Transferase Buffer. The resulting oligo(dT)₁₇ is measured by HPLC.