nicked calf thymus dna richardson c c 1966 in procedures in nucleic acid research cantoni g l and davies d r eds vol 1 pp 212 223 harper and row new york or d at copolymers as primer template in the former it is stated that each assay contained 20 u03bcmol of sodium glycinate ph 9 2 2 u03bcmol of mgcl 2 0 3 u03bcmol of 2 mercaptoethanol 10 nmol of each of the four deoxynucleoside triphosphates datp dctp dgtp and dttp and 40 nmol of nicked calf thynus dna nucleotides plus 20 u03bcl of diluted enzyme in a final volume of 0 30 ml protein concentration was determined both by the lowry method lowry o h rosebrough n j farr a l and randall r j 1951 j biol chem 193 265 275 using bovine serum albumin as the standard and by quantitative amino acid analysis on a durrum d 500 analyzer protein analysis by electrophoresis was performed on 10 acrylamide gels under reducing conditions in the presence of sodium dodecyl sulfate according to the procedure of laemmli laemmli u k 1970 nature 227 680 685 optimizing expression of dna polymerase i growth of atl100 is extremely slow at non inducing temperatures with a doubling time t d of 120 minutes in l broth at 32 u00b0 c this is consistent with the strain making very small colonies after overnight growth on l broth plates surprisingly shifting a broth grown culture of atl100 to the inducing temperature 42 u00b0 c.