Below is a short step-by-step guide demostrating how the NWLSS™ Derivative Application can be used. The example uses data from a typical experiment using the NWLSS™ NWK-MDA01 assay. It may be more convenient to print this page or open it in another browser window (or tab).

Action	Result
•Click the "Worksheet" link.	•The Derivative Spectroscopy Worksheet is displayed. The worksheet provides input boxes for the Savitzky-Golay parameters, data file and data set selection.
•Enter the Savitzky-Golay parameters.	• Polynomial(M): 3   Derivative(d): 3    Filter length(f): 29   Scale(s): 1000000
•Select the MDA data file in the Example files list.	•The radio button is highlighted.
•Press Process.	•The properties for the selected file are displayed in the upper right quadrant of the display. Plots of the Zero Order Spectrum, Convolution Key and Derivative Spectrum are shown in the center of the page. Note: On opening the Worksheet the 3rd-derivative of a Guassian curve is displayed by default. Because the plots scale to the data, the C0-1 ([MDA]=0) data set will appear to have significant noise because the range of derivatives is near zero.
•Select C1-1 in the Select Data Set list.	•Data set C1-1 is the spectra of the 1 µM MDA calibrator.
•Click Display	•The zero order, convolution key (no change) and derivative spectra for sample C1-1 are displayed.
•Move the cursor over the peak in the Zero Order Spectrum plot.	•The xy coordinate of the trace under the cursor is displayed below the plot. The value displayed is persistent until a new coordinate is detected under the cursor. The Zero Order and Derivative plots are independent.
•Move the cursor over the Spectral Data scroll bar.	•A set of xy data pairs are contained in a scrollable list. These are the actual data used to calculate the derivative and plot the zero order spectrum for the data set.
•Click on Download	•The Derivative Spectra for all the data sets contained in the data file are downloaded as a comma separated file (csv). This allows collection of raw data for documentation and further analysis offline. Depending on the user's operating system and browser, this csv file can be opened or saved to the local computer.
•Click on Calibration	•The Calibrator Assignment page will open.
•Enter the calibrator string 0,1,2,3,4 in the Calibrator value list input box and press Assign.	•The Calibrator Assignment Table is displayed listing all data set headers (sample ids) in the current data file. The a radio button is associated with each calibrator value.
•Select the appropriate radio buttons for each calibrator.	•One radio button for each calibration data set is "on". The calibrators can be in any order and replication count.
•Press Calculate	•The Results page opens displaying the calibration curve for the data file and the interpolated analyte concentration.
•Press Download	•The results for the data file calibration on interpolation are downloaded as a csv to the user's local computer. This file is useful for documentation as well as further analysis.