Product Class: Other

Taq DNA Ligase
NEBU recombinant 45 No

Taq DNA Ligase has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10233580.

Try HiFi Taq DNA Ligase, for superior fidelity and thermostability

Product Introduction

Taq DNA Ligase will ligate these substrates:

Nicked DNA/RNA





Catalog # Size Concentration
M0208S 2000.0 units 40000 units/ml
M0208L 10000.0 units 40000 units/ml

Featured Videos

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    DNA Ligation

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    Avoiding RNase Contamination

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    NEBioCalculator® - Using the Ligation module

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    What are the best conditions for DNA ligation?

Product Information

Description

Taq DNA Ligase is a thermostable ligase that catalyzes the formation of a phosphodiester bond between the 5´-phosphate and the 3´-hydroxyl of two adjacent DNA strands. The strands to be ligated need to be hybridized and accurately paired, with no gap, to a complementary DNA strand; allowing resolution of single nucleotide variants. Taq DNA Ligase uses NAD as a cofactor and it is active at elevated temperatures (37° C – 75° C).  

Product Source

Purified from an E. coli strain containing the cloned ligase gene from Thermus thermophilus HB8 (1-3).
This product is related to the following categories:
DNA Ligases Products
This product can be used in the following applications:
Gibson Assembly®,
Cloning Ligation

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

(Cohesive End Unit)
One unit is defined as the amount of enzyme required to give 50% ligation of the 12-base pair cohesive ends of 1 µg of BstEII-digested λ DNA in a total reaction volume of 50 µl in 15 minutes at 45°C.

Reaction Conditions

1X Taq DNA Ligase Reaction Buffer
Incubate at 45°C

1X Taq DNA Ligase Reaction Buffer
20 mM Tris-HCl
25 mM Potassium Acetate
10 mM Magnesium Acetate
1 mM NAD 1
10 mM DTT
0.1% Triton® X-100
(pH 7.3 @ 25°C)

Usage Concentration

40,000 units/ml

Storage Buffer

10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
200 µg/ml Recombinant Albumin
50% Glycerol
pH 7.4 @ 25°C

Heat Inactivation

No

Unit Assay Conditions

1X Taq DNA Ligase Reaction Buffer and DNA (20 µg/ml). After incubation at 45°C for 15 minutes, the reaction is terminated by addition of stop dye (50% glycerol, 50 mM EDTA and bromophenol blue), heated at 70°C for 10 minutes and then loaded on a 0.7% agarose gel. Due to the presence of ligase, the cos ends of BstEII-digested λ DNA will stay together after 70°C heat treatment.

Application Features

  • Allele-specific gene detection using Ligase Detection Reaction and Ligase Chain Reaction (4).
  • Mutagenesis by incorporation of a phosphorylated oligonucleotide during primer extension amplification (5).

Product Notes

  1. Reaction Conditions: Incubate DNA and enzyme in 1X Taq DNA Ligase Buffer at 45°C for 15 minutes or in a thermocycler with a program suited to the reaction described by Barany (1991) Genetic Disease Detection and DNA Amplification Using Cloned Thermostable Ligase. Proc. Natl. Acad. Sci. USA 88, 189-193. The reaction is stopped with a mixture of 50% glycerol, 50 mM EDTA, bromphenol blue.
  2. 1X Taq DNA Ligase Reaction Buffer requires NAD+ as a cofactor. NAD+ is supplied in the 10X Taq DNA Ligase Reaction Buffer; the buffer should be stored at -80°C to extend the half life of the NAD+ cofactor.
  3. Taq DNA ligase will not ligate short 4-base overlaps (typical of restriction enzyme digests), while it efficiently ligates 12-base pair overhangs.

References

  1. Takahashi, M. et al. (1984). J. Biol. Chem.. 259, 10041-10047.
  2. Barany, F. (1991). Proc. Natl. Acad. Sci. USA.. 88, 189-193.
  3. Barany, F. and Gelfand, D.H. (1991). Gene.. 109, 1-11.
  4. Barany, F. (1991). The Ligase Chain Reaction in a PCR World.. 5-16.
  5. Michael, S.F. (1994). Biotechniques.. 16, 411-412.

Protocols, Manuals & Usage

Protocols

  1. Protocol for Taq DNA Ligase (M0208)

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. Is Taq DNA Ligase used for a special technique?
  2. How much ligation occurs at mismatches when using Taq DNA Ligase?
  3. How many temperature cycles will Taq DNA Ligase survive?
  4. Can Taq DNA Ligase be used for cloning?
  5. What is the molecular weight of Taq DNA Ligase?
  6. Why is the Taq DNA Ligase buffer brown?
  7. Does Taq DNA Ligase require NAD?
  8. What is the activity of Taq DNA Ligase in other NEBuffers?
  9. What is the activity of Taq DNA Ligase at various temperatures?
  10. What is the stability of Taq DNA Ligase at 95°C?
  11. What is the stability of Taq DNA Ligase at room temperature?
  12. What is LCR and which enzymes do you recommend?
  13. What is LDR and how does it differ from LCR?
  14. How can I design probes for LDR or LCR to maximize specificity?
  15. Do thermostable DNA ligases (such as Taq DNA Ligase, 9°N DNA Ligase, and HiFi Taq DNA Ligase) ligate sticky ends?

Troubleshooting

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.

Specification Change Notifications

Taq DNA Ligase has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10233580. All subsequent (higher number) lots will contain rAlbumin.

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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