How does qbase+ deal with replicates?
Technical (PCR) replicates
- qbase+ automatically deals with technical replicates or repeated measurements.
- PCR wells with an identical sample and target name are recognized as technical replicates.
At the very beginning of the calculation workflow, the software averages the Cq values of all (non-empty) wells of a replicate group. Outliers can be removed before calculations.
- Custom sample properties defining biological groups can be added as extra information to the samples (Sample properties window). They enable statistical analysis and grouping of results in bar charts.
Inter-run calibrators (IRCs)
- Inter-run calibration is a calculation procedure to detect and remove inter-run variation. The experimenter measures one or more identical samples for the same target in different runs.
These identical samples are called inter-run calibrators (IRCs) and they are used to detect and correct inter-run variation.
- To avoid interpretation of inter-run calibrators as technical replicates, they should have a different sample name in the different runs (e.g. IRC1_a, IRC1_b, ...). Users should then indicate that a number of sample names actually refer to the same biological sample that is used as an inter-run calibrator (e.g. both IRC2_a and IRC2_b refer to the second inter-run calibrator, sample IRC2).
This procedure is known as setting the inter-run calibrators.