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

UniProt accession no.

storage temp.

−20°C

Gene Information

bacteriophage T4 ... 30(1258680)

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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

Certificate of Analysis

Certificate of Origin

Xin Cheng et al.
European journal of cell biology, 91(10), 782-788 (2012-08-04)
Translocation of mitochondrial DNA (mtDNA) fragments to the nucleus and insertion of those fragments into nuclear DNA has been observed in several organisms ranging from yeast to plants and mammals. Disruption of specific nuclear genes by de novo insertions of...
Engler, M.J. and Richardson, C.C. et al.
The Enzymes, 5, 3-3 (1982)
Hiroshi Ochiai et al.
Proceedings of the National Academy of Sciences of the United States of America, 109(27), 10915-10920 (2012-06-20)
To understand complex biological systems, such as the development of multicellular organisms, it is important to characterize the gene expression dynamics. However, there is currently no universal technique for targeted insertion of reporter genes and quantitative imaging in multicellular model...
M J Moore et al.
Science (New York, N.Y.), 256(5059), 992-997 (1992-05-15)
A simple and efficient method for synthesizing long, site-specifically modified RNA molecules was developed whereby segments of RNA were joined with the use of bacteriophage T4 DNA ligase. A single hydrogen or O-methyl group was substituted for the 2'-hydroxyl group...
Wenxin Gu et al.
Bioorganic & medicinal chemistry letters, 22(11), 3693-3698 (2012-05-09)
A series of 2,6-disubstituted aminoalkoxypyrimidine carboxamides (AAPCs) with potent inhibition of bacterial NAD(+)-dependent DNA ligase was discovered through the use of structure-guided design. Two subsites in the NAD(+)-binding pocket were explored to modulate enzyme inhibitory potency: a hydrophobic selectivity region...
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.
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