Taq DNA Ligase

Reagents Supplied

The following reagents are supplied with this product:

NEB #	Component Name	Component #	Stored at (°C)	Amount	Concentration
M0208S     -20       Taq DNA Ligase M0208SVIAL -20 1 x 0.05 ml 40,000 units/ml   Taq DNA Ligase Reaction Buffer B0208SVIAL -20 1 x 1.5 ml 10 X
M0208L     -20       Taq DNA Ligase M0208LVIAL -20 1 x 0.25 ml 40,000 units/ml   Taq DNA Ligase Reaction Buffer B0208SVIAL -20 1 x 1.5 ml 10 X

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).

Companion Products

• Monarch® Plasmid Miniprep Kit 
• Monarch® DNA Gel Extraction Kit
• Monarch® PCR & DNA Cleanup Kit (5 μg)

Materials Sold Separately

• λ DNA-BstEII Digest
• Taq DNA Ligase Reaction Buffer

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.

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.

1. Protocol for Taq DNA Ligase (M0208)

• Tips for Maximizing Ligation Efficiencies

• DNA Ligase Selection Chart
• Properties of DNA and RNA Ligases

• NEBcloner^®
• NEBioCalculator®
• Thermostable Ligase Reaction Temperature Calculator

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 Tips for Ligation Reactions

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 & Change Notifications

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]

• M0208S_L_v2

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]

• M0208S_L_v1_0101507
• M0208S_L_v1_0101602
• M0208S_L_v1_0111608
• M0208S_L_v1_0111702
• M0208S_L_v2_0111708
• M0208S_L_v2_0111803
• M0208L_v2_10013340
• M0208L_v2_10027166
• M0208S_v2_10027168
• M0208L_v2_10037482
• M0208S_v2_10037483
• M0208L_v2_10042318
• M0208L_v2_10045767
• M0208S_v2_10045768
• M0208L_v2_10051891
• M0208L_v2_10055360
• M0208S_v2_10056661
• M0208L_v2_10061276
• M0208S_v2_10061277
• M0208L_v2_10065582
• M0208L_v2_10072387
• M0208S_v2_10065583
• M0208L_v2_10079381
• M0208L_v2_10084266
• M0208S_v2_10084267
• M0208L_v2_10088977
• M0208L_v2_10092256
• M0208L_v2_10099151
• M0208L_v2_10105424
• M0208S_v2_10088976
• M0208L_v2_10111322
• M0208L_v2_10118314
• M0208L_v2_10121462
• M0208S_v2_10111325
• M0208L_v2_10131484
• M0208L_v2_10144150
• M0208L_v2_10149072
• M0208L_v2_10156105
• M0208S_v2_10157013
• M0208L_v2_10168274
• M0208L_v2_10176027
• M0208S_v2_10176795
• M0208L_v2_10178439
• M0208L_v2_10185547

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

• Taq DNA Ligase
• Taq DNA Ligase Reaction Buffer

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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.