Product Class: Other

E. coli DNA Ligase
NEBU recombinant 16 65 Heat

 E.coli DNA Ligase has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10232139.

Product Introduction

E. coli DNA Ligase will ligate these substrates:

dsDNA




Nicked DNA/RNA



Catalog # Size Concentration
M0205S 200.0 units 10000 units/ml
M0205L 1000.0 units 10000 units/ml

Product Information

Description

Highlights

  • Specific Activity: 6,000 units/mg
  • Supplied with 10X Reaction Buffer
E. coli DNA Ligase catalyzes the formation of a phosphodiester bond between the 5´-phosphate and the 3´-hydroxyl of two adjacent DNA strands in duplex DNA with cohesive ends. It is not appreciably active on blunt-ended substrates. E. coli DNA Ligase uses NAD as a cofactor and can be heat-inactivated. E. coli DNA Ligase is active at a range of temperatures (4° C – 37° C). 

Product Source

Purified from E. coli strain containing a cloned E. coli DNA Ligase gene.
This product is related to the following categories:
DNA Ligases Products
This product can be used in the following applications:
Non-Cloning Ligation

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  • M0205S     -20    
      E. coli DNA Ligase M0205SVIAL -20 1 x 0.02 ml 10,000 units/ml
      E. coli DNA Ligase Reaction Buffer B0205SVIAL -20 1 x 1 ml 10 X
  • M0205L     -20    
      E. coli DNA Ligase M0205LVIAL -20 1 x 0.1 ml 10,000 units/ml
      E. coli DNA Ligase Reaction Buffer B0205SVIAL -20 1 x 1 ml 10 X

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of λ DNA (5' DNA termini concentration of 0.12 μM, 300 μg/ml) in a total reaction volume of 20 μl in 30 minutes at 16°C in 1X E. coli DNA Ligase Reaction Buffer.

Reaction Conditions

1X E. coli DNA Ligase Reaction Buffer
Incubate at 16°C

1X E. coli DNA Ligase Reaction Buffer
30 mM Tris-HCl
4 mM MgCl2
26 µM NAD
1 mM DTT
50 µg/ml Recombinant Albumin
(pH 8 @ 25°C)

Usage Concentration

10,000 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

65°C for 20 minutes

Application Features

  • Okayama and Berg cDNA cloning (3)

Product Notes

  1. Requires NAD+ (nicotinamide adenine dinucleotide) as a cofactor, in contrast to other ligases which use rATP.
  2. Ligation of blunt-ended fragments is extremely inefficient. For ligation of blunt-ended fragments we recommend Blunt/TA Ligase Master Mix (NEB #M0367).
  3. Does not ligate RNA to DNA (4).
  4. This enzyme ligates only DNA fragments with cohesive termini.

References

  1. Panasenko, S.M. et al. (1977). Science. 196, 188-189.
  2. Panasenko, S.M. et al. (1978). J. Biol. Chem.. 253, 4590-4592.
  3. Okayama, H. and Berg, P. (1982). Mol. Cell. Biol.. 2, 161-170.
  4. Higgins, N.P. and Cozzarelli, N.R. (1979). R. Wu(Ed.), MethodsEnzymol.. 68, 50-71. New York: Academic Press.

Protocols, Manuals & Usage

Protocols

  1. E. coli DNA Ligase Protocol (M0205)

Usage & Guidelines

Tools & Resources

Selection Charts

Web Tools

FAQs & Troubleshooting

FAQs

  1. How does E. coli DNA Ligase (NEB# M0205) differ from T4 DNA Ligase (NEB# M0202)?
  2. Which ligase should I use?
  3. Will E. coli DNA Ligase work in rCutSmart Buffer?

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

E.coli DNA Ligase has been reformulated with Recombinant Albumin (rAlbumin) beginning with Lot #10232139. 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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