As with Endpoint analysis Kinetic analysis leverages Dazdaq’s transform concept to maximise flexibility and simplicity in assay design.
Kinetic analysis will typically be performed on each well’s plot of readings (Y) against cycle or time (X). Kinetic type analysis can also be performed using different plots such as readings against temperature or temperature against time with any axes orientation.
For each kinetic transform, individual kinetic points can be flagged to be ignored from calculation. Also, a sub-range can be specified so only data points within this area are considered, simplifying analysis of sections of contiguous data.
As with endpoint transforms results from one transform can be used as inputs into other transforms for total flexibility in analysis configuration. For example, IC50 can be determined for each kinetic well by fitting a Four Parameter fit into each well, a standard graph can then be plotted for the resulting IC50’s against specified concentrations to determine concentrations from kinetic plots.
A comprehensive set of transforms are provided for analysis of kinetic data:
Calculate the area under each kinetic chart. As with all kinetic transforms, all points within the well can be analysed, or a sub-range which can be specified numerically or using the drag-bars.
Fit any of the installed curve fit methods to each kinetic well. Fitting kinetic data through a model in this way can be used for smoothing data or for identifying patterns in kinetic data. The calculated coefficients of each fit are then available to other transforms and expressions.
Rate of Change
Differentiate kinetic graphs to identify rates of change. The results of this transform could be used as inputs to another transform, such as maximum peaks or area under peak.
Remove an inherent baseline from each kinetic well using a variety of techniques.
Fluorescent Background Correction
Remove the effect of fluorescent background on temperature, typically required for melting curve type analysis (using Temperature/Fluorescence plots), e.g.
Identify up to 4 peaks in kinetic wells using a variety of techniques.
Area under specific peak
Identify areas under specific peaks.
Calculation of Y from X using interpolation
Determine Y value from a specified X value for each well using interpolation. The X value can be specified numerically or using the drag bar.
Identify where the maximum slope occurs in each well. Use the gradient, X or Y or the mid point as inputs to other transforms.
Other included kinetic transforms
||Fit a straight line through each kinetic plot.
||Find the ratio between two sets of kinetic data.
||Calculate the change in Y for each plot.
|Delta X for Delta Y
||Calculate the change in X for a specified change in Y.
||Calculate Y’s for a specified X.
||Identify the single peak or dip of each plot.
|Area under Peaks
||Determine the area under up to 4 peaks.
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