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D8276

Sigma-Aldrich

DNA-Polymerase I, Klenow-Fragment aus E. coli

buffered aqueous glycerol solution

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About This Item

CAS-Nummer:
9012-90-2
MDL-Nummer:
MFCD00283091
UNSPSC-Code:
12352204
NACRES:
NA.53

Qualität

for molecular biology

Qualitätsniveau

200

Form

buffered aqueous glycerol solution

Mol-Gew.

103 kDa

Konzentration

~3,000 units/mL

UniProt-Hinterlegungsnummer

P00582

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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LIG1

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Piktogramme

Health hazard
GHS08

Signalwort

Danger

H-Sätze

H334

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)

Analysenzertifikate (COA)

Suchen Sie nach Analysenzertifikate (COA), indem Sie die Lot-/Chargennummer des Produkts eingeben. Lot- und Chargennummern sind auf dem Produktetikett hinter den Wörtern ‘Lot’ oder ‘Batch’ (Lot oder Charge) zu finden.

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Sigma-Aldrich

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T4549

Trypsin -Lösung aus Schweinepankreas

DNA polymerase versus DNA binding to the anticancer drug, cis-platin.
Bose, R.N., et al.
Inorgorganica Chimica Acta, 300, 937-937 (2000)
R B Wallace et al.
Science (New York, N.Y.), 209(4463), 1396-1400 (1980-09-19)
Many eukaryotic genes contain intevening sequences, segments of DNA that interrupt the continuity of the gene. They are removed from RNA transcripts of the gene by a process known as splicing. The intervening sequence in a yeast tyrosine transfer RNA
H Klenow et al.
Proceedings of the National Academy of Sciences of the United States of America, 65(1), 168-175 (1970-01-01)
Purification of DNA polymerase from E. coli B has in two cases each time led to the isolation of two separate polymerase activities, enzyme A and enzyme B. Enzyme A was in contrast to enzyme B almost completely devoid of
L M Houdebine
Nucleic acids research, 3(3), 615-630 (1976-03-01)
E.Coli DNA polymerase I (Klenow subfragment) was used for the synthesis of complementary DNA with the mRNAs for rabbit milk proteins as templates. The cDNA formed, contained 200 nucleotides and represented about 20% of the mRNA template. The cDNA was
Second-strand cDNA synthesis: mRNA fragments as primers.
U Gubler
Methods in enzymology, 152, 330-335 (1987-01-01)
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