Stocks of PCNA and dsDNA solutions were mixed to final concentrations of 0.4 mM and 1.1 mM, respectively (1 8 trimer:duplex molar ratio), and incubated at room temperature for 30 min before screening crystallization conditions by the hanging drop vapour diffusion method.
The conditioned eye-blink response incidence was above their spontaneous blink rate (21.8 SD 17.5 blinks per minute, assessed 1 min before and after the experiment): conditioned eye-blink response incidence (mean 23.8%) was quantified in a time interval of ∼ 315 ms per trial (see 'Materials and methods' section).
Results show that most of studied protected columns resist around 120 min before collapsing in fire conditions.
The gradient was kept at 100% B for 0.5 min before returning to starting conditions.
The column was then washed at 100% B for 1 min before returning to starting conditions in 7 min.
To normalise readings, the thermometer was placed standing on a chair/bench outside the pitch for 10 15 min before recording the environmental conditions.
× 15 cm × 2 μm) over a 25-min linear gradient from 4%to50%0% buffer B (80% acetonitrile and 0.08% formic acid in Milli-Q water (v/v)), followed by another 2 min to 90% buffer B. The column was then washed by holding the 90% buffer B for 10 min before returning to initial conditions of 96% buffer A (0.1% formic acid in Milli-Q water (v/v)).
These conditions are held for 15 min before returning to the initial conditions.
The other three treatments, T2, T3 and T4, were done with copper sulphate dosages of 15, 30 and 45 g/ton, respectively, and the pulp conditioned for 5 min before adding the collector.
The gradient elution was used with the following conditions: 5% eluent B maintained for 3 min, followed by a linear increase to 45% of eluent B at 25 min, then a further increase to 90% at 30 min, conditions were held constant for 2 min before being decreased to the initial conditions at 34 min followed by a 6 min isocratic wash at 5% to re-equilibrate the column.
