D2886

Sigma-Aldrich

DNA Ligase from T4-infected Escherichia coli

buffered aqueous glycerol solution

Synonym(s):
Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:
NACRES:
NA.53

Quality Level

grade

for molecular biology

form

buffered aqueous glycerol solution

specific activity

4,000 U/mL

mol wt

68 kDa

storage temp.

−20°C

Gene Information

bacteriophage T4 ... 30(1258680)

Looking for similar products? Visit Product Comparison Guide

Application

Suitable for:
  • Ligation of blunt ended or cohesive DNA fragments
  • Ligation of cloning vector and restriction insert fragments
  • Seal nicks in double stranded DNA and RNA or DNA/RNA hybrids
  • Couple RNA single strands by bridging oligonucleotide adapters

Biochem/physiol Actions

T4 DNA Ligase forms an energy dependent phosphodiester linkage between the termini of adjacent polynucleotides of duplex DNA. The ligation reaction requires ATP as a cofactor. Ligation of blunt-ended fragments requires higher enzyme concentration and can be facilitated by using PEG in the reaction mixture. The enzyme requires a 3′ hydroxyl and 5′ phosphate for ligation. Self-ligation of vector DNA can be prevented by dephosphorylation with alkaline phosphatase. T4 ligase plays an active role in repair of DNA and RNA nicks.

Components

T4 DNA Ligase is supplied in a solution containing 20 mM Tris-HCl (pH 7.5), 50 mM KCl, 1 mM DTT, and 50% (v/v) glycerol.

Unit Definition

One Weiss unit is defined as the amount of enzyme required to catalyze the exchange of 1 nmole of P32 from pyrophosphate into ATP as Norit-absorbable material in 20 minutes at 37°C.

Other Notes

T4 DNA Ligase is inactivated by heating at 65 °C for 10 minutes.

Personal Protective Equipment

dust mask type N95 (US),Eyeshields,Gloves

RIDADR

NONH for all modes of transport

WGK Germany

WGK 3

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Justin L Sparks et al.
Molecular cell, 47(6), 980-986 (2012-08-07)
Ribonucleotides are incorporated into DNA by the replicative DNA polymerases at frequencies of about 2 per kb, which makes them by far the most abundant form of potential DNA damage in the cell. Their removal is essential for restoring a...
Adam B Robertson et al.
The Journal of biological chemistry, 287(39), 32953-32966 (2012-08-01)
The Escherichia coli very short patch (VSP) repair pathway corrects thymidine-guanine mismatches that result from spontaneous hydrolytic deamination damage of 5-methyl cytosine. The VSP repair pathway requires the Vsr endonuclease, DNA polymerase I, a DNA ligase, MutS, and MutL to...
Athena Kantartzis et al.
Cell reports, 2(2), 216-222 (2012-09-04)
Trinucleotide repeat (TNR) expansions are the underlying cause of more than 40 neurodegenerative and neuromuscular diseases, including myotonic dystrophy and Huntington's disease. Although genetic evidence points to errors in DNA replication and/or repair as the cause of these diseases, clear...
Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 1-1 (1989)
P Upcroft et al.
Gene, 51(1), 69-75 (1987-01-01)
We describe a system to generate cDNA or genomic libraries from DNA segments that have blunt termini. Background and rearrangement levels are low, but efficiencies are high and the procedural times very short. T4 ligase in the presence of polyethylene...
Protocols
T4 DNA Ligase catalyzes the formation of a phosphodiester bond between the terminal 5′ phosphate and a 3′ hydroxyl groups of duplex DNA or RNA. The enzyme efficiently joins blunt and cohesive ends and repairs single-stranded nicks in duplex DNA, RNA or DNA/RNA hybrids.
Read More

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Contact Technical Service

Social Media

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon

MilliporeSigma

Research. Development. Production.

We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production.

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Reproduction of any materials from the site is strictly forbidden without permission.