D8276
DNA Polymerase I, Klenow Fragment from Escherichia coli
buffered aqueous glycerol solution
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About This Item
Nom du produit
DNA Polymerase I, Klenow Fragment from Escherichia coli, buffered aqueous glycerol solution
grade
Molecular Biology
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
Quality Level
Gene Information
Escherichia coli K12 ... polA(948356)
Catégories apparentées
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
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.
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.
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.
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Classe de stockage
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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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)
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
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
Yossi Weizmann et al.
Journal of the American Chemical Society, 126(4), 1073-1080 (2004-01-30)
The ultra-sensitive magneto-mechanical detection of DNA, single-base-mismatches in nucleic acids, and the assay of telomerase activity are accomplished by monitoring the magnetically induced deflection of a cantilever functionalized with magnetic beads associated with the biosensing interface. The analyzed M13phi DNA
Contenu apparenté
Instructions
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