STANDARD OPERATING PROCEDURE

Manual Analysis with AIGS SOFTWARE K1.2.70.77

For Software Version K1.2.70.77, please double check the graphs for all the test in the Analysis
Screen.
For some questionable results, manual analysis is needed:

1. Both Graphs N1 and N2 go up but with results “NEG,NEG” “NEG,POS“ ”POS,NEG”

If both graphs N1 (blue) and N2 (yellow) show an obviously increase between cycles 15
to 40, but the software judges at least one gene to be negative, then click “Manual” button,
change the "Threshold" to 200 (the default value is 200)and the "Start Cycle" to 8. Then
click "Analysis" button. If the results change to Pos,Pos, then the results are reported for
Pos,Pos. (Fig.01 and Fig.02).
 2 If both graphs N1 (blue) and N2 (yellow) show a very weak increase between cycles
15 to 40, or you are not sure if there are increase, you can also click ”Manual” button,
change the "Threshold" to 200 (the default value is 200) and the "Start Cycle" to 8. Then
click "Analysis" button. If the result still shows Neg,Neg, it is reported for Neg,Neg.
(Fig.03A & Fig.03B)

Fig. 03A

Fig. 03B
2. If only one graph (blue or yellow) shows an increase between cycles 15 to 40, then
the "threshold" needs to be changed to 1000. Then click "Analysis". If the result
changes to Neg,Neg, then the result is reported to Neg,Neg.

3. If Both Graphs N1 and N2 go up with results “Pos, Pos”

For the results of Pos, Pos, after clicking on the "Analysis" menu and selecting a sample with
the results of Pos, Pos, then proceed by clicking the "Auto" button. The displayed results are
reported as the results of that sample (CT Value).