Description
DNA Polymerase I is a DNA polymerase with 5´→3´ and 3´→5´ exodeoxyribonuclease activities. DNA Polymerase I also incorporates biotinylated nucleotides. Applications—DNase I-dependent nick translation, second-strand synthesis in cDNA cloning, fill-in of 5´ overhangs Source—purified from E. coli expressing the DNA Polymerase I gene on a plasmid Performance and quality testing Endodeoxyribonuclease assay; efficiency of DNase I-dependent nick translation determined. Unit definition One unit incorporates 10 nmol of total deoxyribonucleotide into acid-precipitable material in 30 min at 37°C using template/primer. Unit reaction conditions 50 mM potassium phosphate (pH 7.0), 6.7 mM MgCl2, 1 mM 2-mercaptoethanol, 80 g/mL template/primer, 32 M dTTP, 69 nM [3H]dTTP, and enzyme in 100 L for 30 min at 37°C