D8276
DNA Polymerase I, Klenow Fragment from Escherichia coli
buffered aqueous glycerol solution
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About This Item
Produtos recomendados
grau
for molecular biology
Nível de qualidade
forma
buffered aqueous glycerol solution
peso molecular
103 kDa
concentração
~3,000 units/mL
nº de adesão UniProt
atividade externa
Endonuclease, none detected
Condições de expedição
wet ice
temperatura de armazenamento
−20°C
Informações sobre genes
Escherichia coli K12 ... polA(948356)
Descrição geral
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.
Aplicação
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
Componentes
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) .
Definição da unidade
One unit converts 10 nanomoles of deoxyribonucleoside triphosphates into acid insoluble material in 30 min. at 37 °C.
Reconstituição
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.
Nota de análise
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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Nº do produto
Descrição
Preços
Palavra indicadora
Danger
Frases de perigo
Declarações de precaução
Classificações de perigo
Resp. Sens. 1
Código de classe de armazenamento
10 - Combustible liquids
Classe de risco de água (WGK)
WGK 1
Ponto de fulgor (°F)
Not applicable
Ponto de fulgor (°C)
Not applicable
Equipamento de proteção individual
Eyeshields, Gloves, multi-purpose combination respirator cartridge (US)
Certificados de análise (COA)
Busque Certificados de análise (COA) digitando o Número do Lote do produto. Os números de lote e remessa podem ser encontrados no rótulo de um produto após a palavra “Lot” ou “Batch”.
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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
Second-strand cDNA synthesis: mRNA fragments as primers.
U Gubler
Methods in enzymology, 152, 330-335 (1987-01-01)
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
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