Product Class: Other

Terminal Transferase
NEBU recombinant 37 75 Heat

Product Introduction

Terminal transferase catalyzes the addition of deoxynucleotides to the 3' hydroxyl terminus of DNA molecules.

  • Useful for Labeling of the 3′ ends of DNA with modified nucleotides (e.g., ddNTP, DIG-dUTP)
  • Isolated from a recombinant source
  • Supplied with 10X Reaction Buffer
  • Tested for the absence of endonucleases and exonucleases
Catalog # Size Concentration
M0315S 500.0 units 20000 units/ml
M0315L 2500.0 units 20000 units/ml

Product Information

Description

Terminal transferase (TdT) is a template independent polymerase that catalyzes the addition of deoxynucleotides to the 3' hydroxyl terminus of DNA molecules. Protruding, recessed or blunt-ended double or single-stranded DNA molecules serve as a substrate for TdT. The 58.3 kDa enzyme does not have 5' or 3' exonuclease activity. The addition of Co2+ in the reacton makes tailing more efficient.

Product Source

An E. coli strain that carries the cloned Terminal Transferase gene from calf thymus.
This product is related to the following categories:
DNA Labeling Products,
DNA Manipulation Products,
This product can be used in the following applications:
A-tailing,
PCR

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0315S     -20    
      Terminal Transferase M0315SVIAL -20 1 x 0.025 ml 20,000 units/ml
      Terminal Transferase Reaction Buffer Pack B0315SVIAL -20 1 x 1.5 ml 10 X
      CoCl2 B0252SVIAL -20 1 x 1.4 ml 10 X
  • M0315L     -20    
      Terminal Transferase M0315LVIAL -20 1 x 0.125 ml 20,000 units/ml
      Terminal Transferase Reaction Buffer Pack B0315SVIAL -20 1 x 1.5 ml 10 X
      CoCl2 B0252SVIAL -20 1 x 1.4 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme catalyzing the incorporation of 1 nmol dTTP into acid-insoluble material in a total reaction volume of 50ul in 1 hour at 37°C using d(A)18 as primer.

Reaction Conditions

1X Terminal Transferase Reaction Buffer Pack
Supplement with 0.25 mM CoCl2
Incubate at 37°C

1X Terminal Transferase Reaction Buffer Pack
50 mM Potassium Acetate
20 mM Tris-acetate
10 mM Magnesium Acetate
(pH 7.9 @ 25°C)

Storage Buffer

50 mM KPO4
100 mM NaCl
1.43 mM β-ME
50% Glycerol
0.1% Triton® X-100
pH 7.3 @ 25°C

Heat Inactivation

75°C for 20 minutes

Molecular Weight

Theoretical: 58000 daltons

5' - 3' Exonuclease

No

3' - 5' Exonuclease

No

Strand Displacement

No

Unit Assay Conditions

1X Terminal Transferase Reaction Buffer, 0.72 μM d(A)18, 0.2 mM dTTP, and 1 μCi [3H]- dTTP in a 50 μl total reaction volume.

Application Features

  • Addition of homopolymer tails to the 3' ends of DNA
  • Labeling the 3' ends of DNA with modified nucleotides (e.g., ddNTP, DIG-dUTP)
  • TUNEL assay (in situ localization of apoptosis)
  • TdT dependent PCR

References

  1. Chang, L.M. and Bollum, F.J. (1986). CRC Crit. Rev. Biochem.. 21, 27-52.
  2. Roychoudhury, R., Jay, E. and Wu, R. (1976). Nucl. Acids Res.. 3, 101-116.
  3. Tu, C.P. and Cohen, S.N. (1980). Gene. 10, 177-183.
  4. Boule, J.B., Rougeon, F. and Papanicolaou, C. (2001). J. Biol. Chem. 276, 31388-31393.

Protocols, Manuals & Usage

Protocols

  1. A Typical DNA Tailing Reaction
  2. A-Tailing (single nucleotide) with Terminal Transferase

FAQs & Troubleshooting

FAQs

  1. Can Terminal Transferase be heat inactivated?
  2. Can Terminal Transferase label the 5' end of DNA?
  3. Is Terminal Transferase supplied with dNTPs?
  4. Does Terminal Transferase require Co2+ as a divalent cation?
  5. Can just one dNTP be added to the end of DNA with Terminal Transferase?
  6. Does Terminal Transferase have a preference for one type of 3' DNA end?
  7. What are the kM values for Terminal Transferase?
  8. Will Terminal Transferase add a non-radioactively labeled dNTP?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

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