The determination of BamHI steady-state kinetic
parameters using break light B. (a) The observed change
in fluorescence intensity over time of an assay containing a constant
3.2 nM break light B at 37°C (6 mM Tris⋅HCl/100 mM
NaCl/6 mM MgCl2/1 mM DTT, pH 7.5;
λEx = 485 nm, λEm = 517 nM),
BamHI (10 units), and varying nonlabeled substrate
oligonucleotide. Total substrate concentrations (including break
light): 389 nM (○), 196 nM (□), 81 nM (⋄), 42
nM (▵), 11 nM (●), 7.5 nM
(■), and 3.4 nM (⧫). (b) Lineweaver–Burke
plot from a after correction for the carrier dilution
effect.