Terminal Deoxynucleotidyl Transferase

EU Commodity Code

38220090

Shipping Temp

Blue Ice

Storage Temp

-20°C

Recombinant calf thymus TdT|Supplied with Reaction Buffer

TdT is a template-independent DNA polymerase located in the cell nucleus which catalyses irreversible terminal addition, via polymerization, of deoxynucleotides at the 3' hydroxyl ends of oligo or polydeoxynucleotide initiators accompanied by the release of inorganic phosphate.

Applications

• Oligo and DNA 3’-terminal labeling • Homopolymeric tailing of linear duplex DNA with any type of termini • TUNEL assay (in situ localization of apoptosis) • 5’-RACE

Components

• Terminal Deoxynucleotidyl Transferase: 1x500U or 1x2500U • Reaction Buffer (5X): 1x0.4ml or 2x1ml 1M potassium cacodylate, pH 7.2 (25°C), 5mM CoCl2, 0.5mM DTT, 0.05% Triton X-100

Storage Buffer

100mM potassium acetate, pH 6.8, 2mM 2-mercaptoethanol, 0.01% Triton X-100, 50% glycerol.

Unit Definition

1 unit of the enzyme iincorporates 1 nanomole of deoxythymidylate into a polynucleotide fraction in 60 minutes at 37ºC.

Note: Due to the presence of CoCl2, the TdT Reaction Buffer is incompatible with downstream applications. It is necessary to remove CoCl2 from the reaction mixture by spin column or phenol/chloroform extraction and subsequent ethanol precipitation.

Quality Control

Tested for absence of exodeoxyribonucleases, endodeoxyribonucleases ribonucleases, and phosphatases.

Inhibitors

Metal chelators, ammonium chloride, iodide and phosphate ions.

Inactivators

Inactivated by heating at 70°C for 10 mins. or by addition of EDTA.

Storage and Stability

Store at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Source

E. coli cells carrying a cloned gene encoding calf thymus terminal deoxynucleotidyl transferase.

Concentration

~20U/ul

Important Note

This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

References

1. Bollum, F.J., Terminal deoxynucleotidyl transferase, The Enzymes, the third edition (Boyer, P.D., ed.), Academic Press, New York, vol.10, 145-171, 1974.|2. Deng, G.R., Wu, R., Terminal transferase: Use in the tailing of DNA and for in vitro mutagenesis, Meth. Enzymol., 100, 96-116, 1983.|3. Eschenfeldt, W.H., et al., Homopolymeric tailing, Meth. Enzymol., 152, 337-342, 1987.|4. Tu, C.-P.D., Cohen, S.N., 3’-end labeling of DNA with [alfa-32P]cordycepin-5’-triphosphate, Gene, 10, 177-183, 1980.|5. Vincent, C., et al., Synthesis of 8-(2,4-dinitrophenyl-2,6-aminohexyl)aminoadenosine-5’-triphosphate: Biological properties and potential uses, Nucleic Acids Res., 10, 6787-6796, 1982.|6. Kumar, A., et al., Nonradioactive labeling of synthetic oligonucleotide probes with terminal deoxynucleotidyl transferase, Anal. Biochem., 169, 376-382, 1988.|7. Gaastra, W., Klemm, P., Radiolabeling of DNA with terminal transferase, Methods in Molecular Biology, vol.2: Nucleic Acids (Walker, J.M., ed.), Humana, Clifton, NJ, 269-271, 1984.

USBio References

Source: E. coli cells carrying a cloned gene encoding calf thymus terminal deoxynucleotidyl transferase.|Concentration: ~20U/ul||Important Note: This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.

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T2750 Terminal Deoxynucleotidyl Transferase (TdT)

EU Commodity Code

Shipping Temp

Storage Temp

Recombinant calf thymus TdT|Supplied with Reaction Buffer

References

USBio References

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