protein

DNA Ligase T4, 5u/ul

MBS654009

200 Units

360 USD

Enzyme

DNA Ligase T4, 5u/ul

[DNA Ligase T4, 5u/ul]

Molecular Biology Grade

May be stored at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawingand prior to removing the cap.

CAS Number: 9015-85-4. Host Note: E. coli cells carrying a cloned gene 30 from bacteriophage T4. Unit Definition: One Weiss unit of the enzyme catalyzes the conversion of 1nmol of [32PPi] into Norit-adsorbable form in 20 min. at 37°C (1). One Weiss unit is equivalent to approximately 200 cohesive end ligation units (CEU)*. Enzyme activity is assayed in the following mixture: 66mM Tris-HCl, pH 7.6, 6,6mM MgCl2, 0.066mM ATP, 10mM DTT, 3.3uM [32PPi]. *One CEU is defined as the amount of enzyme required to give 50% ligation of HindIII fragments of lambda DNA in 30 min. at 16°C. Buffer: Supplied in 20mM Tris-HCl pH 7.5, 50mM KCl, 1mM DTT, 0.1mM EDTA, 50% glycerol.

Supplied with: MBS654009 DNA Ligase T4 Reaction Buffer, 10X: 400mM Tris-HCl, 100mM MgCl2, 100mM DTT, 5mM ATP, pH 7.8 . PEG Solution (50% Polyethylene glycol 4000). Inhibition and Inactivation: • T4 DNA Ligase is strongly inhibited by NaCl or KCl at concentrations higher than 200mM. • Inactivated by heating at 65°C for 10 min or at 70°C for 5 min. Quality Control: Endodeoxyribonuclease Assay: No conversion of covalently closed circular DNA to nicked DNA was detected after incubation of 50 Weiss units of T4 DNA Ligase with 1ug of pUC19 DNA for 4hours at 37°C. Ribonuclease Assay: No contaminating RNase activity was detected after incubation of 50 Weiss units of T4 DNA Ligase with 1ug of [3H]-RNA for 4 hours at 37°C. Labeled Oligonucleotide (LO) Assay: No degradation of single-stranded and double-stranded labeled oligonucleotide was observed after incubation with 10u of T4 DNA Ligase for 4 hours at 37°C . Blue/White Cloning Assay: The B/W assay was replaced with LO test after validating experiments showed LO test ability to detect nuclease and phosphatase activities with sensitivity that equals to that of B/W test.

Cloning; Cloning-Enzymes

T4 DNA Ligase catalyzes the formation of a phosphodiester bond between juxtaposed 5'-phosphate and 3'-hydroxyl termini in duplex DNA or RNA. The enzyme repairs single-strand nicks in duplex DNA, RNA or DNA/RNA hybrids, joins DNA fragments with either cohesive or blunt termini (3,4). The T4 DNA Ligase requires ATP as cofactor.

~55.3kD

200 Units

360 USD

1000 Units

635