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Table 9. Cycling protocol for Rotor-Gene Q
Step
Time
Temperatur
e
Comment
Initial PCR activation
step
5 min
95°C
Activation of HotStarTaq
Plus
DNA Polymerase
3-step cycling
Data collection at 60°C
for green, crimson,
orange and yellow
channels
Denaturation
15 s
95°C
Annealing
*
15 s
60°C
*
Extension
10 s
72°C
Number of cycles
40
*
Gain optimization before first acquisition at 60°C for green, crimson,
orange and yellow channels
8.
Click “OK” to close the window and return to the Wizard.
9.
To set the gain optimization settings for the Green, Yellow, and Orange
Channels click “Gain Optimisation”.
10.
Select the 3 channels in the drop-down menu and click “Add”.
11.
In the dialog box that opens, confirm the standard settings. Click
“Perform Optimisation Before 1st Acquisition”. Then close the window.
Make sure that the tube at position 1 is not empty, since the gain
optimization will be performed on this tube.
OMAMAN-36 C: Method User Guide/Package Insert
ERP Use Only
January 2017
AOAC Research Institute
Expert Review Panel Use Only