产品名称：Klenow Fragment, exo–
Storage Condition -20 C
10X Reaction Buffer 500 mM Tris-HCl (pH8.0 at 25°C), 50mM MgCl 2 , 10mM DTT.
Definition of Activity Unit
One unit of the enzyme catalyzes the incorporation of 10nmol of deoxyribonucleotides into a polynucleotide fraction (adsorbed on DE-81) in 30min at 37°C, using poly(dA-dT)·poly(dA-dT) as a template·primer.
Enzyme activity is assayed in the following mixture: 67mM potassium phosphate (pH7.4), 6.7mM MgCl 2 , 1mM 2-mercaptoethanol, 0.033mM dATP, 0.033mM dTTP, 0.4MBq/mL [ 3 H]-dTTP, and 62.5µg/mL poly(dA-dT)·poly(dA-dT).
Inactivation Inactivated by heating at 75°C for 10min or by the addition of EDTA.
Inhibition Inhibitors: metal chelators, PP i , P i (at high concentrations) ( see Reference 7).
Molecular Weight 68kDa monomer.
Lacks 3→5 exonuclease activity
Incorporates modified nucleotides (e.g., Cy3-, Cy5-, fluorescein-, rhodamine-, aminoallyl-, biotin-labeled nucleotides)
Active in restriction enzyme, PCR, and RT buffers
Random-primed DNA labeling ( see ? References 2-4)
Labeling by fill-in 5-overhangs of dsDNA
Strand displacement amplification (SDA) ( see ? Reference 5)
DNA sequencing by the Sanger method ( see ? Reference 6)