D2886

DNA Ligase from T4-infected Escherichia coli

buffered aqueous glycerol solution

• Synonyme(s) : Polydeoxyribonucleotide Synthase, Polynucleotide Ligase, T4 DNA Ligase
• Numéro CAS: 9015-85-4
• Numéro de classification (Commission des enzymes): 6.5.1.1 (BRENDA, IUBMB)
• Numéro CE : 232-770-0
• Numéro MDL: MFCD00163508
• Code UNSPSC : 12352204
• Nomenclature NACRES : NA.53

Propriétés

• Qualité: for molecular biology
• Niveau de qualité: 200
• Forme: buffered aqueous glycerol solution
• Activité spécifique: 4,000 U/mL
• Poids mol.: 68 kDa
• Numéro d'accès UniProt: P00970
• Température de stockage: −20°C
• Informations sur le gène: bacteriophage T4 ... 30(1258680)

Description

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

Actions biochimiques/physiologiques

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.

Composants

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.

Définition de l'unité

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.

Autres remarques

T4 DNA Ligase is inactivated by heating at 65 °C for 10 minutes.

Informations sur la sécurité

• Pictogrammes: GHS08
• Mention d'avertissement: Danger
• Mentions de danger: H334
• Conseils de prudence: P261 - P284 - P501
• Classification des risques: Resp. Sens. 1
• Code de la classe de stockage: 10 - Combustible liquids
• Classe de danger pour l'eau (WGK): WGK 1
• Point d'éclair (°F): Not applicable
• Point d'éclair (°C): Not applicable
• Équipement de protection individuelle: Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)