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D8276

Sigma-Aldrich

DNA Polymerase I, Klenow Fragment from Escherichia coli

buffered aqueous glycerol solution

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About This Item

Numero CAS:
Numero MDL:
Codice UNSPSC:
12352204
NACRES:
NA.53

Grado

for molecular biology

Forma fisica

buffered aqueous glycerol solution

PM

103 kDa

Concentrazione

~3,000 units/mL

N° accesso UniProt

Attività estranea

Endonuclease, none detected

Condizioni di spedizione

wet ice

Temperatura di conservazione

−20°C

Informazioni sul gene

Escherichia coli K12 ... polA(948356)

Descrizione generale

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.

Applicazioni

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

Componenti

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) .

Definizione di unità

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

Ricostituzione

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.

Risultati analitici

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.

Pittogrammi

Health hazard

Avvertenze

Danger

Indicazioni di pericolo

Consigli di prudenza

Classi di pericolo

Resp. Sens. 1

Codice della classe di stoccaggio

10 - Combustible liquids

Classe di pericolosità dell'acqua (WGK)

WGK 1

Punto d’infiammabilità (°F)

Not applicable

Punto d’infiammabilità (°C)

Not applicable

Dispositivi di protezione individuale

Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)


Certificati d'analisi (COA)

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Proceedings of the National Academy of Sciences of the United States of America, 65(1), 168-175 (1970-01-01)
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