DNA Polymerase I, Klenow Fragment from Escherichia coli, buffered aqueous glycerol solution

Descriptions

Analysis Note

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.

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

Components

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) .

General description

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.

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.

Unit Definition

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

Specifications

No detailed specifications are available for this product.

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Code	Description	Quality Level	Manufacturer	Form	Price		Quantity
3577272	DNA Polymerase I, Klenow Fragment from Escherichia coli, buffered aqueous glycerol solution	200	SIGMA-ALDRICH	buffered aqueous glycerol solution	£345.43 (exc VAT) per EACH		- +
3577273	DNA Polymerase I, Klenow Fragment from Escherichia coli, buffered aqueous glycerol solution	200	SIGMA-ALDRICH	buffered aqueous glycerol solution	£182.46 (exc VAT) per EACH		- +