D8276

DNA Polymerase I, Klenow Fragment from Escherichia coli

buffered aqueous glycerol solution

• Numéro CAS: 9012-90-2
• Numéro MDL: MFCD00283091
• Code UNSPSC : 12352204
• Nomenclature NACRES : NA.53

Propriétés

• Qualité: for molecular biology
• Forme: buffered aqueous glycerol solution
• Poids mol.: 103 kDa
• Concentration: ~3,000 units/mL
• Numéro d'accès UniProt: P00582
• Activité étrangère: Endonuclease, none detected
• Conditions d'expédition: wet ice
• Température de stockage: −20°C
• Informations sur le gène: Escherichia coli K12 ... polA(948356)

Description

Description générale

DNA polymerase I yields two fragments (small and large) upon protease digestion. The large fragment (Klenow fragment) loses the 5′ exonuclease activity that is present in the intact holoenzyme. However, it retains both the polymerase 5′→3′ activity and the 3′→5′ exonuclease activity of the native enzyme.

Application

Suitable for:

• DNA sequencing by the Sanger dideoxy method
• Synthesis of the complementary strand of cDNA
• Filling in 5′-overhangs in double stranded DNA to form blunt ends
• Mutagenesis of DNA with second strand synthesis using oligonucleotides
• Labeling DNA by the random primer method

Composants

DNA Polymerase I is supplied as a solution in 50 mM Tris-HCl (pH 7.5), 1 mM EDTA, 5 mM dithiothretol, and 50% glycerol (v/v) .

Définition de l'unité

One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min. at 37 °C.

Reconstitution

The enzyme solution may be diluted with 50 mM Tris-HCl, pH 7.5, 100 mM ammonium sulfate, 10 mM 2-mercaptoethanol, and 1 mg/ml bovine serum albumin.

Remarque sur l'analyse

The activity is assayed in a reaction mixture containing 50 mM potassium phosphate (pH 7.5), 3 mM MgCl₂, 1 mM 2-mercaptoethanol, 32.5 μM ³²P-dATP, 32.5 μM dTTP, 62.5 μg/ml poly(dA-dT) and 0.01-1 unit enzyme.

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)