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You are here:Home » Molecular Biology » Cloning-Enzymes » DNA Polymerase I Klenow Fragment

DNA Polymerase I Klenow Fragment

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Specifications

DNA Polymerase I Large Fragment exhibits the 5’ ---> 3’ polymerase and 3’ ---> 5’ exonuclease activity of DNA Polymerase. Lacks 5’ ---> 3’ exonuclease activity.
Catalog #D3940
Applications• Random primer labeling
• Sanger sequencing
• cDNA Second strand synthesis
• 3’ end-labeling, filling
Storage and StabilityAliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
CAS Number9012-90-2
Molecular Weight62kD
SourceE.coli CM 5199
PreparationPrepared by subtilisin proteolysis of DNA Polymerase I, retaining the 5’ -3’ polymerase and the 3’-5’ exonuclease cayalytic activities of the native enzyme but lacks the 5’ -3’ exonuclease activity.
Storage Buffer (Form)Supplied as a liquid in 50mM potassium phosphate, pH 7.0 with 1mM DTT, 50% glycerol.
Concentration~10u/ul
Unit Definition1u catalyzes the incorporation of 10nmoles of deoxynucleotides into polynucleotide fraction using poly(dA-dT)•poly(dA-dT) as template•primer in 30 minutes at 37°C.
Quality Control
Activity Assay
67mM potassium phosphate, pH 7.4, 6.7mM magnesium chloride, 1mM 2-mercaptoethanol, 0.033mM dATP, 0.033mM dTTP, 0.4MBq/ml [H3]-dTTP, 62.5ug/ml poly(dA-dT)•poly(dA-dT).
Endodeoxyribonuclease Assay
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 20u of Klenow Fragment with 1ug of pBR322 DNA in 50ul of Reaction Buffer for 4hours at 37°C.
Function Assay
Klenow Fragment was tested for filling-in recessed 3’-termini of DNA.
Exonuclease, Endonuclease, RibonucleaseNone detected
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.
Alternate namesE. coli lysogenic CM 5199


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