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

DNA Polymerase I

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Specifications

DNA Polymerase I catalyzes the polymerization of nucleotides into duplex DNA in the 5’=>3’ direction. The enzyme exhibits 3’-5’ proofreading exonuclease activity and 5’=>3’ degrading activity. Purified from an E.coli strain carrying a DNA polymerase I overproducing plasmid.
Catalog #D3935
Application• Second strand cDNA synthesis
• Nick translation
Supplied withD3935-05: DNA Polymerase I Reaction Buffer, 10X
Storage and StabilityMay be stored at 4°C for short-term only. Aliquot 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.
Molecular Weight109kD
Source
E. coli, strain carrying a DNA polymerase I overproducing plasmid.
Form
Supplied as a liquid in PBS, pH 7.5, 1mM DTT, 50% glycerol.
Concentration
10u/ul
D3935-05, Reaction Buffer (10X)Supplied as a liquid in 500mM Tris-HCl, pH 7.5 (25°C), 100mM MgCl2, 10mM DTT
Unit Definition
One unit of enzyme will catalyze the exchange of 10nmole of total DNA to a DE-81 adsorbable form in 30 minutes at 37°C using poly[dA-dT)] as a template primer.
Quality Control
Endodeoyuribonuclease Assay
No detectable conversion of covalently closed circular DNA to nicked DNA was observed after incubation of 20u of DNA Ploymerase with 1ug of pBR322 DNA in 50ul of reaction buffer for 4 hours at 37°C.
Activity Assay
67mM potassium phosphate (pH 7.4), 6.7mM MgCl2, 1mM 2-mercaptoethanol, 0.033mM dATP, 0.033mM dTTP, 0.4MBq/ml [3H]-dTTP and 62.5ug/ml poly(dA-dT)•poly(dA-dT).
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 namesEC=2.7.7.7; E. coli DNA Pol I Supplied with 10X Reaction Buffer


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