D2886
DNA Ligase from T4-infected Escherichia coli
buffered aqueous glycerol solution
Synonym(s):
Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase
grade
Molecular Biology
Quality Level
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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Related Categories
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.
Other Notes
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.
T4 DNA Ligase is inactivated by heating at 65 °C for 10 minutes.
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.
Signal Word
Danger
Hazard Statements
Precautionary Statements
Hazard Classifications
Resp. Sens. 1
Storage Class Code
10 - Combustible liquids
WGK
WGK 1
Flash Point(F)
Not applicable
Flash Point(C)
Not applicable
Personal Protective Equipment
dust mask type N95 (US), Eyeshields, Gloves
Regulatory Information
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Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 1-1 (1989)
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
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
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
Ayako Nishizawa-Yokoi et al.
The New phytologist, 196(4), 1048-1059 (2012-10-12)
Evidence for the involvement of the nonhomologous end joining (NHEJ) pathway in Agrobacterium-mediated transferred DNA (T-DNA) integration into the genome of the model plant Arabidopsis remains inconclusive. Having established a rapid and highly efficient Agrobacterium-mediated transformation system in rice (Oryza
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