D8276
DNA-Polymerase I, Klenow-Fragment aus E. coli
buffered aqueous glycerol solution
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About This Item
Empfohlene Produkte
Qualität
Molecular Biology
for molecular biology
Form
buffered aqueous glycerol solution
Mol-Gew.
103 kDa
Konzentration
~3,000 units/mL
UniProt-Hinterlegungsnummer
Fremdaktivität
Endonuclease, none detected
Versandbedingung
wet ice
Lagertemp.
−20°C
Angaben zum Gen
Escherichia coli K12 ... polA(948356)
Allgemeine Beschreibung
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.
Anwendung
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
Komponenten
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) .
Einheitendefinition
One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min. at 37 °C.
Rekonstituierung
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.
Hinweis zur 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.
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Danger
H-Sätze
P-Sätze
Gefahreneinstufungen
Resp. Sens. 1
Lagerklassenschlüssel
10 - Combustible liquids
WGK
WGK 1
Flammpunkt (°F)
Not applicable
Flammpunkt (°C)
Not applicable
Persönliche Schutzausrüstung
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
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Sambrook, J.F., et al.
Molecular Cloning: A Laboratory Manual, 5-5 (1989)
R B Wallace et al.
Science (New York, N.Y.), 209(4463), 1396-1400 (1980-09-19)
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Second-strand cDNA synthesis: mRNA fragments as primers.
U Gubler
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