Since each step of a flow based assay is best performed at a flow rate, which may be different from other operations on the same flow path, an important component for optimization of reagent based assays is flow programming (Introduction and Chapter 1)
Therefore SI protocol for reagent-based assays uses the following combination of flow rates:
- fast flow rates for filling and flushing the system with carrier solution (up to 200 μL/sec)
- lintermediate flow rates for sample and reagent injection (10 to 50 μL/sec) and metering of flow reversal volume
- moderate flow rates for analyte desorption from microcolumn (5μL/sec)
- low flow rates for analyte adsorption on a column (0.5 to 5 μL/sec)
- stopped flow for slow reactions or reaction rate measurement in the flow cell
Selection of Flowrate
2.1.3.
