1.

PROGRAMMING VIRAL LOAD TESTING

Click Absolute The interface for setting up a new absolute quantification experiment opens

1.1. Set up Detectors

Click Detector

Enter the
- Experiment properties,
- Experiment name (is generated automatically but can be modified),
- User name.
- Add comments to the experiment.

Enter the name of the target and choose the reporter dye.
If you are using a Human Real-Time PCR Assay (HIV-1, HCV, HBV, CMV), choose FAM.

Click Add Detector to add another target. Enter the name of the target and choose the reporter dye. If you are
using a Human Real-Time PCR Assay (HIV-1, HCV, HBV, CMV), choose HEX for the Internal Control (IC).

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 1.2. Set up the sample information

Click Sample, enter the sample ID and press Enter to add the sample to the sample ID list.
If you want to perform batch testing Batch Add can be chosen.
- Enter the start sample ID and the number of samples,
- Click Add and samples will be added automatically.

Sample Name can be entered by clicking in the respective field. Further fields can be added by the user.

1.3. Set up the plate

Click Plate
- Select a well on the plate for your sample
- Select the corresponding sample on the left side
- Select the assay item and set up the properties of your sample (Unknown, Standard, Non-template Control
and Positive Control).
- Add the corresponding concentration (Conc.) of each standard (provided in the handbook of the assay kit).
Select the Concentration Units. For Human Real-Time PCR assays select IU/ml.

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 1.4. Set up the program

Click Program and a default temperature profile is opened.

To modify the temperature profile, the following modifications can be used:

Add Stage
- Hold Stage
- PCR Stage

Add Step
- Number of Cycles
- Liquid Quantity of the Test
- Temperature of a step
- Time of a step
- Measuring Step

Delete

Click in respective field to enter desired temperature and duration of each step. Measuring is to be check
marked during elongation step (72 °C).

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An example of a modified cycling profile is shown.

The set up can be saved as template file.

Click  Save Save as Template

The file will be stored in the template folder.

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Open the HumaCycler.
- Ensure that water-condensate seal ring is put on the block
- Place the in accordance with the assay kit handbook prepared samples/tubes on the reaction block.
- Close the tray (snaps when it is locked).

Click on Run and the Run Status display will be shown.

- To start the run click on Start Run.

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Confirmation window will pop up. Confirm by clicking OK.

Fluorescence, temperature curves and the program can be observed during the run by choosing the
corresponding item. Furthermore, the run status is shown, e.g. start and end time, current cycle and step.

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