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T4 DNA Ligase for Ligation of dsDNA, NGS library prep and molecular cloning

T4 DNA Ligase

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T4 DNA Ligase is an ATP and Mg2+ dependent dsDNA ligase which catalyses the formation of a phosphodiester bond between 3’-hydroxyl and 5’-phosphate termini in duplex DNA, duplex RNA and some DNA/RNA hybrids. T4 DNA Ligase is active on both blunt-end and cohesive-end substrates. It is also completely inactivated by incubating at 70°C for 10 minutes.

This is a high-quality (commercial grade) version of the T4 DNA Ligase. T4 DNA Ligase is recombinantly produced in E. coli. ArcticZymes’ T4 DNA Ligase is extensively tested for contaminating DNase and RNase activities as well as residual host-cell gDNA.

Key Features

  • ATP and Mg2+ dependent dsDNA ligase
  • Easily heat-inactivated at 70°C for 10 minutes
  • Extensively tested for contaminating DNase and RNase activities as well
    as residual host-cell gDNA

Applications

  • Ligation of dsDNA
  • NGS library prep
  • Molecular cloning

Figures

Properties

Specifications
Unit Definition

0.1 units is defined as the amount of enzyme that is needed to convert 1 pmol (of 18 pmol) of nicked DNA substrate in 20 minutes at 25 °C in a 20 µl reaction volume in a buffer consisting of 62.5 mM Tris-HCl pH 7.5, 10 mM DTT, 1 mM ATP, 0.05 mg/ml BSA and 25 mM KCl.

One Weiss Unit is equivalent to approximately 500 ArcticZymes Units.

Size

56.9 kDa

Inactivation

T4 DNA Ligase can be completely inactivated by incubating at 70°C for 10 minutes.

Storage & Stability

The enzyme is stable at -20°C for > 1 year in the supplied storage buffer.The enzyme tolerates at least four freeze-thaw cycles (-80°C) without loss of activity.

Specifications

ArcticZymes is dedicated to the quality of our products. T4 DNA Ligase is manufactured at our ISO 13485 certified facility in Norway.

dsDNA endonuclease activity

10 000 U T4 DNA Ligase was incubated with a supercoiled plasmid (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any transformation of closed circular DNA to nicked DNA.

sDNA endonuclease activity

10 000 U T4 DNA Ligase was incubated with M13 ssDNA (0.5 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of ssDNA degradation.

Exonuclease activity

10 000 U T4 DNA Ligase was incubated with either 3H-dATP labelled ds or ssDNA (0.5 µg, 500 bp) for 4 hours at 37°C. Acid soluble radioactivity from labelled DNA was not significantly over blank test for either substrate.

RNase activity

5000 U T4 DNA Ligase was incubated with a 2 kb RNA transcript (1 µg) for 4 hours at 37°C. Agarose gel electrophoresis did not reveal any visible signs of RNA degradation.

E. coli gDNA contamination

5000 U T4 DNA Ligase was analysed in a probe-based qPCR assay detecting the 23S ribosomal subunit in E. coli. No E. coli gDNA could be detected (LOD: < 3 E. coli genomic copies).

Ordering Info

Quantity
Product Name
T4 DNA Ligase
T4 DNA Ligase
Catalog Nr.
71800-202
71800-100
Pack Size
250 kU
as per request
Concentration
5000
as per request
Price
Add To Cart

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