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Salt-T4® DNA Ligase

Catalog # Concentration Size List Price Quantity Your Price
M0467L 400000 units/ml 100000 units $418.00
$376.20
M0467S 400000 units/ml 20000 units $104.00
$93.60
Catalog # Size List Price Your Price
M0467L 100000 units $418.00
$376.20
M0467S 20000 units $104.00
$93.60
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

Salt-T4 will ligate these substrates:

dsDNA




Nicked DNA/RNA





 


Featured Videos
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  • Monarch_MiniprepProtocol_VideoThumb
    DNA Ligation
  • Monarch_MiniprepProtocol_VideoThumb
    Quick Tips - Do I need a 5' phosphate for ligation?
  • Monarch_MiniprepProtocol_VideoThumb
    NEBioCalculator® - Using the Ligation module
  • Monarch_MiniprepProtocol_VideoThumb
    Quick Tip - What is the ideal incubation temperature for ligation?
  • Monarch_MiniprepProtocol_VideoThumb
    Quick Tips - Troubleshooting problematic ligation reactions
  • Monarch_MiniprepProtocol_VideoThumb
    Quick Tips - How do I calculate how much DNA to add to a ligation reaction?
For details on NEB's quality controls for DNA ligases, visit our Ligase Quality page. 

A salt tolerant variant of T4 DNA Ligase, Salt-T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5´ phosphate and 3´ hydroxyl termini in duplex DNA or RNA and is designed to function at higher salt concentrations than wild type T4 DNA Ligase. This enzyme will join cohesive end termini at salt concentrations as high as 300 mM without any loss in activity. This enzyme is insensitive to salt carried over from other reaction components (vector or insert DNA) and allows ligation reactions to proceed in alternative reaction buffers with higher levels of salt (e.g., NEBuffer 3.1).

Figure 1: Salt-T4 DNA Ligase displays increased salt tolerance on a cohesive end substrate as compared to T4 DNA Ligase.



400 units of T4 DNA Ligase (NEB #M0202) or Salt-T4 DNA Ligase (NEB #M0467) was incubated with 0.12 μM HindIII (cohesive end) fluorescent-labeled fragments at 25°C for 10 minutes in the presence of 1X T4 DNA Ligase Buffer containing increasing amounts of salt (0-1000 mM NaCl). Ligation products were detected by capillary electrophoresis. 
T4 DNA Ligase rapidly loses activity at salt concentrations above 200 mM, whereas Salt-T4 DNA Ligase retains > 60% activity on a cohesive end substrate at salt concentrations as high as 500 mM. 

Product Source
An E. coli strain that carries a plasmid encoding the engineered Salt-T4 DNA Ligase gene.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  Salt-T4® DNA Ligase M0467SVIAL -20 1 x 0.05 ml 400,000 units/ml
  T4 DNA Ligase Reaction Buffer B0202SVIAL -20 1 x 1 ml 10 X
  StickTogether™ DNA Ligase Buffer B0535AVIAL -20 1 x 1 ml 2 X
  NaCl B5019AVIAL -20 1 x 1 ml 1,000 mM
  Salt-T4® DNA Ligase M0467LVIAL -20 1 x 0.25 ml 400,000 units/ml
  T4 DNA Ligase Reaction Buffer B0202SVIAL -20 1 x 1 ml 10 X
  StickTogether™ DNA Ligase Buffer B0535AVIAL -20 3 x 1 ml 2 X
  NaCl B5019AVIAL -20 1 x 1 ml 1,000 mM
Features
  • Cloning of restriction fragments
  • Joining linkers and adapters to blunt-ended DNA
An exception occurred during the operation, making the result invalid. Check InnerException for exception details.

Companion Products
Materials Sold Separately
Notes
  • ATP is an essential cofactor for the reaction. This contrasts with E. coli DNA ligase which requires NAD.
  • To dilute Salt-T4 DNA Ligase that will subsequently be stored at -20°C, 50% glycerol storage buffer (Diluent Buffer A, NEB #B8001S) should be used; to dilute for immediate use, 1X T4 DNA Ligase Reaction Buffer can be used.

  • Ligation Temperature

    For convenience, ligations may be done at room temperature (20-25°C). For cohesive (sticky) ends (1X T4 DNA Ligase Buffer), use 1 µl of Salt-T4 DNA Ligase in a 20 µl reaction for 10 minutes. For blunt ends or single base overhangs (1X StickTogether™ DNA Ligase Buffer), use 1 µl of Salt-T4 DNA Ligase in a 20 µl reaction for 10 minutes. Do not heat kill the reaction because heat treating the PEG in the StickTogether™ DNA Ligase Buffer will inhibit transformation.

References
  • Engler, M.J. and Richardson, C.C. (1982). P.D. Boyer(Ed.), 5, San Diego: Academic Press.
Quality Control Assay
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.
Change Notifications
Effective June 16, 2022, buffer volume will change to 1.0 ml.
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 Data Sheets
Legal And Disclaimer

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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