Bio-Rad BioLogic DuoFlow Pathfinder 80 System DuoFlow Chromatography System Starter Kit - Manual (Page 31)

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Two-Point Correction (best for gradient applications)
a. In the manual screen, set the pH to 8.1, the salt composition to 0 %B and
the flow rate to 2.0 ml/min.
b. Take the column out of line, if it has been connected, and start the pump.
c. When the pH has stabilized, read the pH from the status bar or, alternatively,
collect the effluent and measure the pH using a high quality Tris compatible
pH probe.
d. Change %B to 100 % (or to the maximum %B that will be used for the
experiment).
e. When the pH reading has stabilized, read the pH from the status bar or,
alternatively, collect the effluent and measure the pH using a high quality Tris
compatible pH probe.
f. Set the %B back to 0% and re-equilibrate the system with the low salt buffer.
g. From the buffer blending setup screen, place a check in the Use pH
Correction box.
h. Set the desired pH to 8.1 and the Observed at 0 %B pH to the pH measured
in step (c).
i. Place a check in the Use Two Point Correction box and set %B to the value
used in step (d)).
j. Set the Observed at %B pH to the pH measured in step (e).
k. Press OK.
Column Equilibration.
a. Connect the UNO Q1 column.
b. Wash the column with 6.5 ml (5 column volumes) of pH 8.10 Tris (100% B) at
2 ml/min.
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