D2886
DNA Ligase from T4-infected Escherichia coli
buffered aqueous glycerol solution
Synonym(s):
Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase
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About This Item
Recommended Products
grade
for 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.
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.
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Description
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Glutathione S-Transferase from E. coli, recombinant, expressed in E. coli, buffered aqueous solution
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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
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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
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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
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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