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D2886

Sigma-Aldrich

DNA Ligase from T4-infected Escherichia coli

buffered aqueous glycerol solution

Sinónimos:

Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase

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About This Item

Número de CAS:
9015-85-4
Comisión internacional de enzimas:
6.5.1.1 (BRENDA, IUBMB)
EC Number:
232-770-0
MDL number:
MFCD00163508
UNSPSC Code:
12352204
NACRES:
NA.53

grade

for molecular biology

Quality Level

200

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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Referencia del producto
Descripción
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P4978

Phosphatase, Alkaline from calf intestine, buffered aqueous glycerol solution

P4390

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G5663

Glutathione S-Transferase from E. coli, recombinant, expressed in E. coli, buffered aqueous solution

D7291

Deoxyribonuclease I RNase-free solution from bovine pancreas

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Ribonucleasa A from bovine pancreas, for molecular biology, ≥70 Kunitz units/mg protein, lyophilized

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P6911

Proteasa from Streptomyces griseus, BioReagent, DNase, RNase, and nickase, none detected (No RNase.)

LIG2

QuickLink™ DNA Ligation Kit, joins blunt-end and sticky-end DNA fragments

LIG1

DNA Ligation Kit

pictograms

Health hazard
GHS08

signalword

Danger

hcodes

H334

Hazard Classifications

Resp. Sens. 1

Storage Class

10 - Combustible liquids

wgk_germany

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)

Certificados de análisis (COA)

Busque Certificados de análisis (COA) introduciendo el número de lote del producto. Los números de lote se encuentran en la etiqueta del producto después de las palabras «Lot» o «Batch»

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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
Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 1-1 (1989)
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
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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