One unit is defined as the amount of enzyme required to catalyze the incorporation of 10 nmol of dNTP into acid-soluble form in 30 minutes at 72°C in the presence of the reaction buffer.
Functional assay
Human genomic DNA was amplified using the DNA Polymerase and specific primers to produce a distinct band of 1100 bp.
Self-priming activity
Standard PCR is carried out without primers, using the DNA Polymerase and human genomic DNA. No products were amplified.
Exonuclease assay
Linearized lambda/HindIII fragments are incubated with the DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.
Endonuclease assay
lambda DNA is incubated with the DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.
Nick Activity
Supercoiled plasmid DNA is incubated with the DNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No conversion of covalently closed circular DNA to nicked DNA was detected.
Contamination with E. coli DNA
A sample of the denatured DNA Polymerase is analyzed with specific primers targeting the 16S rRNA gene in qPCR for the presence of contaminating E. coli DNA. No E. coli was detectable.
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