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Merck

D2886

DNA Ligase from T4-infected Escherichia coli

buffered aqueous glycerol solution

Sinónimos:

Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase

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Número CAS:
UNSPSC Code:
12352204
NACRES:
NA.53
EC Number:
232-770-0
MDL number:
Número CE:
Specific activity:
4,000 U/mL
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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)

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.


pictograms

Health hazard

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

Clase de almacenamiento

10 - Combustible liquids

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

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



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Science (New York, N.Y.), 256(5059), 992-997 (1992-05-15)
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Engler, M.J. and Richardson, C.C. et al.
The Enzymes, 5, 3-3 (1982)
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