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Cat. No.: RT4DL-40k (for 40KU)

Cat. No.: RT4DL-200k (for 200KU)

Description

Rapid T4 DNA Ligase is an upgraded version of T4 DNA Ligase, capable of rapidly and efficiently catalyzing the formation of phosphodiester bonds between the 5’-P and 3’-OH ends of double-stranded DNA or RNA with either sticky or blunt ends. This catalytic reaction requires ATP as a cofactor. Additionally, Instant T4 DNA Ligase can repair single-strand nicks in double-stranded DNA, double-stranded RNA, or DNA/RNA hybrids.

Applications

Rapid T4 DNA Ligase is commonly used for the ligation of DNA fragments with vectors, linkers, or adaptors. It can also be used for nick repair and ligase-mediated RNA detection.

Source 

Rapid T4 DNA Ligase is expressed in Escherichia coli, with the expression gene being a mutant of the T4 bacteriophage T4 DNA Ligase gene.

Activity Definition

One unit is defined as the amount of enzyme required to achieve 50% ligation of HindIII fragments of λ DNA in 30 minutes at 16°C in a 20µl assay mixture containing 50mM Tris, pH 7.5, 10mM MgCl₂, 10mM DTT, 1mM ATP, 25µg/ml BSA, and a 5’-DNA termini concentration of 0.12µM (300µg/ml). 200U equals one Weiss unit. The product is available in packages of 200U and 1000U, measured in Weiss units.

Purity

Free from DNA endonucleases, exonucleases, and phosphatases, and free from RNases, meeting the requirements for standard ligation reactions.

Enzyme Storage Solution

20mM Tris, pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, and 50% (v/v) glycerol.

10X Ligation Buffer

400mM Tris, pH 7.8, 100mM MgCl₂, 100mM DTT, 10mM ATP.

Inactivation or Inhibition

Incubation at 65°C for 10 minutes can inactivate Instant T4 DNA Ligase; concentrations of NaCl or KCl greater than 200mM strongly inhibit Instant T4 DNA Ligase.

Storage

Store at -20°C, effective for at least one year.

Precautions

• For routine transformation of Escherichia coli, purification of the ligation product is not necessary; the ligation product can be used directly for transformation. However, for electroporation transformation of Escherichia coli, it is generally recommended to purify the DNA using a DNA purification kit or phenol-chloroform extraction before electroporation.
• For rapid ligation, it is recommended to use Instant DNA Ligation Kit.
• Routine ligation reactions do not require gel electrophoresis observation. If gel electrophoresis observation of the ligation product is needed, it is recommended to incubate at 65°C for 10 minutes to inactivate Instant T4 DNA Ligase, to avoid band shift caused by the binding of Instant T4 DNA Ligase to DNA.
• This product is for scientific research use only by professionals and is not intended for clinical diagnosis or treatment, food, or drug use, and should not be stored 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
• 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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SBS Genetech is a long-term sponsor of Cold Spring Harbor Laboratory