Hi-T4™ DNA Ligase
Product informationCode | Name | Size | Quantity | Price | |
---|---|---|---|---|---|
M2622S |
Hi-T4 DNA Ligase |
20000 units ( 400000 units/ml ) | - | Unavailable in your region | |
M2622L |
Hi-T4 DNA Ligase |
100000 units ( 400000 units/ml ) | - | Unavailable in your region |
Hi-T4™ DNA Ligase
Catalog #M2622
Product Introduction
Hi-T4 DNA Ligase will ligate these substrates:
dsDNA
Nicked DNA/RNA
- Hi-T4 DNA Ligase is an engineered variant of T4 DNA Ligase with improved thermostability compared to T4 DNA Ligase
- Capable of joining blunt end and cohesive end termini, as well as repairing single stranded nicks in duplex DNA and some DNA/RNA hybrids (1)
- Supplied with both T4 DNA Ligase Buffer and StickTogether™ DNA Ligase Buffer
- Other T4 DNA Ligase products include Quick Ligation Kit, Salt-T4, Instant Sticky-end Ligase Master Mix and Blunt/TA Ligase Master Mix
- This product can be purchased in larger volumes. Submit an inquiry to find out more.
- Not sure which ligase to choose? Refer to our DNA and RNA Ligase Properties Chart or DNA Ligase Selection Chart
- Product Information
- Protocols, Manuals & Usage
- Tools & Resources
- FAQs & Troubleshooting
- Citations & Technical Literature
- Quality, Safety & Legal
- Other Products You May Be Interested In
Product Information
Description
Highlights
- Engineered thermotolerant variant of T4 DNA Ligase
- Isolated from a recombinant source
A thermotolerant variant of T4 DNA Ligase, Hi-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 temperatures than wild type T4 DNA Ligase. Hi-T4 DNA Ligase will join blunt end and cohesive end termini as well as repair single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids at temperatures as high as 50ºC.
Product Source
An E. coli strain that carries a plasmid encoding the engineered Hi-T4 DNA Ligase gene.- This product is related to the following categories:
- DNA Ligases Products
- This product can be used in the following applications:
- BioBrick® Assembly,
- Cloning Ligation
Properties & Usage
Unit Definition
One unit is defined as the amount of enzyme required to give 50% ligation of 6 µg of Lambda-HindIII DNA in 30 minutes at 25°C in a total reaction volume of 20 µl.Reaction Conditions
1X T4 DNA Ligase Reaction Buffer
Incubate at 25°C
1X T4 DNA Ligase Reaction Buffer
50 mM Tris-HCl
10 mM MgCl2
1 mM ATP
10 mM DTT
(pH 7.5 @ 25°C)
Storage Buffer
10 mM Tris-HCl
50 mM KCl
1 mM DTT
0.1 mM EDTA
50% Glycerol
pH 7.4 @ 25°C
Heat Inactivation
65°C for 10 minutesFeatures
- Cloning of restriction fragments
- Joining linkers and adapters to blunt-ended DNA
Related Products
Companion Products
- Quick Blunting™ Kit
- Monarch® Plasmid Miniprep Kit
- Monarch® DNA Gel Extraction Kit
- Monarch® PCR & DNA Cleanup Kit (5 μg)
- T4 DNA Ligase
- Nuclease-free Water
Materials Sold Separately
Product Notes
- ATP is an essential cofactor for the reaction. This contrasts with E. coli DNA ligase which requires NAD.
- To dilute Hi-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 cohesive (sticky) ends (1X T4 DNA Ligase Buffer), incubate 1 µl of Hi-T4 DNA Ligase in a 20 µl reaction between 25-50ºC for 10 minutes. For blunt ends or single base overhangs (1X StickTogether DNA Ligase Buffer), incubate 1 µl of Hi-T4 DNA Ligase in a 20 µl reaction between 25-50ºC 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.
Protocols, Manuals & Usage
Protocols
Usage & Guidelines
Tools & Resources
Selection Charts
Web Tools
FAQs & Troubleshooting
FAQs
- What are some causes of ligation reaction failure that can lead to transformation failure?
- When should Hi-T4 DNA Ligase be the enzyme of choice?
- Which buffer should I use with Hi-T4 DNA Ligase?
- What is the activity of Hi-T4 DNA Ligase in other buffers?
- Can this ligase be heat inactivated?
- What is the difference between the unit definitions of T4 DNA Ligase (NEB #M0202) and Hi-T4 DNA Ligase (NEB #M2622)?
- How much DNA should be used in a ligation using this ligase?
Citations & Technical Literature
Citations
Additional Citations
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.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]- M2622L_v1_10058735
- M2622S_v1_10058734
- M2622S_v1_10062171
- M2622L_v1_10065552
- M2622L_v1_10075404
- M2622S_v1_10075408
- M2622S_v1_10092876
- M2622L_v1_10097783
- M2622S_v1_10115457
- M2622L_v1_10111329
- M2622L_v1_10138794
- M2622S_v1_10138795
- M2622L_v1_10150755
- M2622L_v2_10162037
- M2622S_v2_10162038
- M2622L_v2_10180905
- M2622S_v2_10180904
- M2622L_v3_10192174
- M2622S_v3_10208064
- M2622S_v3_10225779
- M2622S_v3_10240891
- M2622L_v3_10250261
- M2622S_v3_10251871
- M2622L_v3_10256904
- M2622S_v3_10256905
- M2622L_v3_10267518
Safety DataSheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.Hi-T4™ DNA Ligase
T4 DNA Ligase Reaction Buffer
StickTogether™ DNA Ligase Buffer
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.
Other Products You May Be Interested In
The supporting documents available for this product can be downloaded below.