E. coli DNA Ligase

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

Application Features

• Okayama and Berg cDNA cloning (3)

Companion Products

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

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.

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.

1. E. coli DNA Ligase Protocol (M0205)

• Activity of DNA Modifying Enzymes in rCutSmart™ Buffer
• Tips for Maximizing Ligation Efficiencies

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

• NEBcloner^®
• NEBioCalculator®

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 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]

• M0205S_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]

• M0205S_L_v2_0911703
• M0205S_L_v2_0911711
• M0205S_L_v2_0911804
• M0205S_v2_10008136
• M0205S_v2_10028144
• M0205L_v2_10028143
• M0205L_v2_10030914
• M0205S_v2_10025798
• M0205S_v2_10045747
• M0205L_v2_10044925
• M0205L_v2_10053290
• M0205L_v2_10054228
• M0205S_v2_10065800
• M0205L_v2_10065801
• M0205L_v2_10094095
• M0205S_v2_10094102
• M0205L_v2_10102812
• M0205S_v2_10123941
• M0205L_v2_10132241
• M0205S_v2_10146340
• M0205L_v2_10146341
• M0205L_v2_10168546
• M0205S_v2_10168545

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

• E. coli DNA Ligase

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 [email protected].

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.