Bio-Rad Experion Protein Analysis Kits User Manual
Page 20
Fig. 1.9. Results table generated for well 2 in Test 1.
6.
To perform a qualitative evaluation of quantitation reproducibility, overlay the
electropherograms of replicate samples. For example, in the virtual gel, select lane 1
and hold the Shift key while selecting lane 12. The electropherograms from all 12 wells
overlap (Figure 1.10). Tighter overlaps signify better reproducibility.
Fig. 1.10. Electropherogram overlay showing the overlay of all 12 of the 100 ng/µl samples.
7.
Evaluate the reproducibility of quantitation by calculating the coefficient of variation
(%CV), which is defined as the standard deviation/mean x 100. To do this, export the
data to a Microsoft Excel or other spreadsheet application either by entering the RNA
concentration values manually or by using the data export feature of Experion software.
Reproducibility is most strongly influenced by pipetting technique (the ability to
reproducibly pipet 1 µl sample and 5 µl loading buffer into each well).
8.
To use the export feature of Experion software, select File > Export Data... The Export
dialog opens. Select Result Table and Format as Single Table and then click Export.
In the Save As window, name the file and designate a folder for it. Click Save.
9.
Launch Excel software and select File > Open. Change Files of type to All, locate the
text file exported in step 8, and select Open. The Import wizard opens. Click Finish.
16
10010511A.qxp 2/28/2008 7:14 AM Page 16