DNA Polymerase I

Detailní popis

Thermo Scientific DNA Polymerase I, a template-dependent DNA polymerase, catalyzes 5'→3' synthesis of DNA. The enzyme also exhibits 3'→5' exonuclease (proofreading) activity, 5'→3' exonuclease activity, and ribonuclease H activity.

Highlights

• Incorporates modified nucleotides (e.g. biotin-, digoxigenin-, aminoallyl-, fluorescently-labeled nucleotides)
• Active in multiple buffers, including restriction enzyme, PCR, and RT buffers

Applications

• DNA labeling by nick-translation in conjunction with DNase (see References 1-3)
• Second-strand synthesis of cDNA in conjunction with RNase H (see Reference 4)

  10X Reaction Buffer	500 mM Tris-HCl (pH 7.5 at 25°C), 100 mM MgCl2, 10 mM DTT.
  CategoryName
  Definition of Activity Unit	One unit of the enzyme catalyzes the incorporation of 10 nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30 min at 37°C, using poly(dA-dT)·poly(dA-dT) as a template·primer. Enzyme activity is assayed in the following mixture: 67 mM potassium phosphate (pH 7.4), 6.7 mM MgCl2, 1 mM 2-mercaptoethanol, 0.033 mM dATP, 0.033 mM dTTP, 0.4 MBq/mL [3H]-dTTP, and 62.5 µg/mL poly(dA-dT)·poly(dA-dT).
  Hazardous	No
  Hazardous:	No
  Inactivation	Inactivated by heating at 75°C for 10 min or by addition of EDTA.
  Inhibition	Inhibitors: metal chelators, PPi, Pi (at high concentrations) (see Reference 5).
  Molecular Weight	103 kDa monomer
  Quality Control	The absence of endodeoxyribonucleases confirmed by appropriate quality test.
  Shelf Life:
  Shipping Condition:
  Shipping Information
  Source	E.coli cells with a cloned polA gene.
  SpecificationName
  SpecificationValue
  Storage Buffer	The enzyme is supplied in 25 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, and 50% (v/v) glycerol.
  Storage Condition	-20 C
  Storage Condition: