DNA Ligase T4, 5u/ul Enzyme
DNA Ligase T4, 5u/ul
Purity
Molecular Biology Grade
Synonyms
DNA Ligase T4; 5u/ul; 5u/ul enzyme
Purity/Purification
Molecular Biology Grade
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.
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)
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.
• 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.
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.
Preparation and Storage
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.
Related Product Information for DNA Ligase T4, 5u/ul enzyme
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.
Product Categories/Family for DNA Ligase T4, 5u/ul enzyme
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Product Notes
The DNA Ligase T4, 5u/ul (Catalog #AAA654009) is an Enzyme and is intended for research purposes only. The product is available for immediate purchase. It is sometimes possible for the material contained within the vial of "DNA Ligase T4, 5u/ul, Enzyme" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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