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Merck

D8276

DNA Polymerase I, Klenow Fragment from Escherichia coli

buffered aqueous glycerol solution

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CAS 번호:
UNSPSC Code:
12352204
NACRES:
NA.53
MDL number:
Concentration:
~3,000 units/mL
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grade

Molecular Biology

Quality Level

form

buffered aqueous glycerol solution

mol wt

103 kDa

concentration

~3,000 units/mL

UniProt accession no.

foreign activity

Endonuclease, none detected

shipped in

wet ice

storage temp.

−20°C

Gene Information

Escherichia coli K12 ... polA(948356)

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.

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

Preparation Note

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.

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.

Other Notes

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


pictograms

Health hazard

signalword

Danger

hcodes

Hazard Classifications

Resp. Sens. 1

저장 등급

10 - Combustible liquids

wgk

WGK 1

flash_point_f

Not applicable

flash_point_c

Not applicable

ppe

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



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관련 콘텐츠


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)
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
DNA polymerase versus DNA binding to the anticancer drug, cis-platin.
Bose, R.N., et al.
Inorgorganica Chimica Acta, 300, 937-937 (2000)



국제 무역 품목 번호

SKUGTIN
D8276-100UN04061833651551
D8276-50UN04061833589359