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Cat. No.: T7DL-150k (for 150KU)

Cat. No.: T7DL-600k (for 600KU)

Cat. No.: T7DL-3m (for 3000KU)

Description

T7 DNA Ligase is an ATP-dependent double-stranded DNA ligase derived from T7 bacteriophage, and it has a much higher efficiency for ligating sticky ends compared to blunt ends.

Unlike T3 and T4 DNA Ligase, T7 DNA Ligase can only catalyze the formation of phosphodiester bonds between adjacent 5’ phosphate and 3’ hydroxyl groups of double-stranded DNA with sticky ends, but it cannot effectively catalyze the ligation of blunt-ended double-stranded DNA. Adding 20% or more PEG6000 can appropriately enhance the blunt-end DNA ligation activity of this enzyme. Therefore, in molecular biology experiments where both blunt-ended and sticky-ended double-stranded DNA substrates are present, and only the sticky-ended double-stranded DNA needs to be ligated, T7 DNA Ligase is an ideal choice.

Applications

Cloning of DNA fragments cut by restriction endonucleases, ligation of double-stranded DNA and adapters, circularization of linear double-stranded DNA, nick repair of double-stranded DNA, site-directed mutagenesis, Golden Gate Assembly of DNA fragments for Transcription Activator-Like Effector Nucleases (TALEN), and sticky-end specific ligation.

Source

Purified from recombinant E. coli strains carrying the gene encoding T7 bacteriophage DNA ligase.

Activity Definition

One unit is defined as the amount of enzyme required to achieve 50% ligation of 100 ng HindIII fragments of λ DNA in a total reaction volume of 20 μl in 30 minutes at 25°C in 1X T7 DNA Ligase Reaction Buffer.

Purity

Free from other types of DNA ligases, DNA endonucleases, exonucleases, RNases, and phosphatases.

Inactivation or Inhibition

Incubate at 65°C for 10 minutes in a reaction system without PEG6000.

Storage

Store at -20°C, valid for two years.

Precautions

• ATP is an essential cofactor for the catalytic activity of T7 DNA Ligase, unlike E. coli DNA Ligase which uses NAD as a cofactor.
• T7 DNA Ligase cannot effectively catalyze the ligation of blunt-ended double-stranded DNA fragments. For blunt-ended double-stranded DNA ligation, it is recommended to use T4 DNA Ligase.
• The substrate for T7 DNA Ligase is double-stranded DNA; it cannot be used for ligation reactions involving single-stranded DNA or RNA.
• The reaction system for T7 DNA Ligase contains 7.5% PEG6000. If PEG6000 cannot be added to the experimental system, consider preparing a ligation reaction buffer without PEG6000, or using the ligation buffer system for T4 DNA Ligase. However, the activity of T7 DNA Ligase in the T4 DNA Ligase buffer system will be reduced by approximately tenfold.
• This product is intended for scientific research use by professionals only and is not for clinical diagnosis or treatment, food or drug use, or storage in ordinary residential areas.
• For your safety and health, please wear a lab coat and disposable gloves when handling.

DNA Ligase Products:

• T4 DNA Ligase
• T4 DNA Ligase (glycerol-free)
• Rapid T4 DNA Ligase
• T4 DNA Ligase (Fast)
• T7 DNA Ligase
• E. coli DNA Ligase
• PBCV-1 DNA Ligase
• T3 DNA Ligase
• Taq DNA Ligase
• Pfu DNA Ligase
• High-Fidelity DNA Ligase
• High-Salt DNA Ligase

SBS Genetech is recognized as one of the global major leading industry players in Ligase Market by third-party market researchers. For more details, please visit Ligase Market - A Global and Regional Analysis Focus on Product, Source, Application, End User, and Country - Analysis and Forecast, 2022-2032

Only for research and not intended for treatment of humans or animals

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