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D2886

Sigma-Aldrich

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

CAS Number:
9015-85-4
Enzyme Commission number:
6.5.1.1 (BRENDA, IUBMB)
EC Number:
232-770-0
MDL number:
MFCD00163508
UNSPSC Code:
12352204
NACRES:
NA.53

grade

for molecular biology

form

buffered aqueous glycerol solution

specific activity

4,000 U/mL

mol wt

68 kDa

UniProt accession no.

P00970

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.

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Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334

Precautionary Statements

P261 - P284 - P501

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

Certificates of Analysis (COA)

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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
Tiansheng Wang et al.
Bioorganic & medicinal chemistry letters, 22(11), 3699-3703 (2012-05-09)
A series of 4-amino-pyrido[2,3-d]pyrimidin-5(8H)-ones were designed and synthesized as a novel class of inhibitors of NAD(+)-dependent DNA ligase that possess potency against Gram-positive bacteria.
Tara Sugrue et al.
Stem cells (Dayton, Ohio), 31(1), 137-145 (2012-09-11)
The regeneration of the hematopoietic system following total body irradiation is supported by host-derived mesenchymal stromal cells (MSCs) within the bone marrow. The mechanisms used by MSCs to survive radiation doses that are lethal to the hematopoietic system are poorly
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
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
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Protocols

T4 DNA Ligase

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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