Product Instructions

Introduction

This user guide provides essential information for operating the Dualo 32® R2 instrument. It covers everything from initial setup and instrument connection to data analysis for your qPCR experiments.

1. SETUP

Installing your Dualo 32 R2

Upon receiving your Dualo 32 R2, you should find the following items in the flight case:

1. Dualo 32 R2 qPCR Instrument
2. Power Supply Unit and Mains Cable
3. Ethernet Cable
4. USB Drive

Retain the flight case and outer box for future storage or shipment protection.

Lid

The lid ensures correct tube seating, provides optical isolation, features a heated element to reduce condensation, and prevents dust from entering empty wells.

USB Port

Use the USB port to insert a USB drive for running experiments.

Display LEDs

The instrument uses display LEDs to indicate its operational status:

Dualo 32 R2 Connections

The Dualo 32 R2 has three connection ports: one on the front and two on the back.

? Power: Connects the instrument to the provided power supply unit.

? Ethernet Port: Connects the instrument to your LAN or computer.

Powering on your Dualo 32 R2

Place the instrument on your lab bench and connect AC power. After a few seconds, the instrument will turn on. The display LEDs will initially be blue, then turn green if the lid is closed, or red if it is open. The Dualo 32 R2 is now ready to run. It uses a 3-pin IEC mains connector; ensure a grounded supply or provide an additional ground connection if necessary.

Connecting your Dualo 32 R2 to a Network, PC or Laptop

Connect one end of the Ethernet cable to the instrument and the other end to a LAN port or directly to your computer. Once connected to a LAN, you can also connect wirelessly via Wi-Fi if your network supports it.

Dualo 32 R2 Heated Lid

The heated lid can become hot during operation; avoid touching it. It preheats to 105 °C upon detecting user activity to expedite run starts. After 5 minutes of inactivity, it turns off to conserve energy.

⚠️ CAUTION: HEATED LID - DO NOT TOUCH

Installing your Dualo 32 R2 Software

For Windows (Mac OS X is also available): Double-click the Dualo 32 R2-Windows-Installer and follow the on-screen instructions.

2. TIPS

Here are some useful tips for operating your Dualo 32 R2 instrument:

1. Lid gets warm: The Dualo 32 R2 lid can get warm during operation; this is normal.
2. Do not leave the lid open: Leaving the lid open can allow dust to enter the wells, affecting instrument performance.
3. Spin your tubes: This ensures all well contents are at the bottom, improving performance.
4. Remove all bubbles: Bubbles can cause optical artefacts. Ensure no bubbles are present in reaction volumes.
5. Keep your lab clean: Maintain a clean workspace, including lab equipment, surfaces, pipettes, and tube racks, to ensure instrument cleanliness and good results.
6. Keep your instrument clean: Perform routine cleaning as described in the Maintenance section.
7. PC settings: Disable all power-saving settings (e.g., sleep, hibernate) on your PC.

3. EXPERIMENT

This section guides you through setting up and running experiments on the Dualo 32 R2. You will learn to create, save, open, and close experiments, set up sample information, and run new experiments via the software or a USB drive.

Choose a New Experiment

The Dualo 32 R2 software displays the home screen. Click 'New' to start a new experiment.

Select Template File

Choose a template to use. You can also add or remove template files. Highlighting a template and clicking 'Select' applies pre-set parameters like temperature profile, optical, and analysis settings.

Click 'Add' to include a new template file.

Experiment Summary

After selecting a template, the Experiment Summary screen appears. The default experiment name includes a date and time stamp, which can be edited. This summary provides details about the instrument, chosen settings, and any notes you add.

Open an Experiment

Click 'Open' to load an existing experiment file. These files contain data and analyses but cannot be re-run.

USB Run

To open a USB run, navigate to the USB location in the 'Open' dialog, select 'Experiment File (.ppf)' in the 'Files of Type' dropdown, and double-click the desired run file.

Saving an Experiment

Experiments can be saved automatically to a pre-set location or manually. Saving before closing is crucial to prevent data loss.

Automatic Saving

When starting a run, you can configure auto-save settings. Select a directory and ensure the 'Automatically save runs on completion' checkbox is ticked. You can choose not to see this dialog again.

Manual Saving

After a run, click 'Save' or 'Save As' to manually save the experiment. 'Save' overwrites the current file, while 'Save As' allows choosing a new location and filename.

Closing an Experiment

Select 'Close'. If the experiment has been edited and not saved, a dialog prompts you to save changes before closing.

Samples

Samples or targets can be defined before or after a run starts. They can be added or removed from the experiment using the '+' and '-' buttons in the 'Samples' pane. The order of samples can be adjusted using the Up and Down arrows.

Assigning Samples to Wells

Assign samples to wells by selecting the wells and clicking 'Set'. Unassign by selecting wells and clicking 'Clear'.

Assigning Targets to Wells

Assign targets as 'Standard' (✅ Std.), 'Unknown' (❓ Unk.), or 'Negative' (❌ Neg.). Clear assignments using the 'Clear' button. Contact technical support for more information on standard controls.

Starting a Run from the Software

Select 'Start Run'. You will be prompted with auto-save options and then asked to choose an instrument from the 'Registered Instruments' list. Press 'Select' to begin the run.

Starting a Run from a USB Drive

Select 'Start Run', then choose 'Start run from USB'. Navigate to your USB drive, press 'Save', and safely remove the drive. Inserting the USB into the instrument will start the experiment automatically.

Loading Strips into your Dualo 32 R2

To ensure the heated lid is balanced, the mount must contain at least one strip in rows A and D, or single tubes in wells A1, A8, D1, and D8. These positions can hold reagents or be empty.

4. ANALYSIS

Pre-set analysis types are available depending on the chosen template.

Auto Quantification

Click 'Auto. Quant.' in the Analysis pane on the left side of the screen to perform auto quantification.

Quantifiers

If you have assigned standards with known quantifiers (e.g., a dilution series), you can view reaction efficiency by selecting 'Quantifiers'. This displays the relationship between Cq and input template quantity, along with derived statistics.

Auto Tm Calling (Melting)

Select the 'Automatic Tm Calling' analysis to identify and characterize melting peaks. Results are displayed below.

5. EXPORT

After analyzing your data, you can export results in various formats, from raw data to custom reports.

Data Export

Export data from result panes in tabular format by clicking the 'Export...' button.

Available formats:

• CSV: Editable data file compatible with spreadsheet applications.
• PDF: Suitable for archiving, printing, and presentations; not editable.

Report Generation

Generate customizable reports by selecting 'Experiment Report' under the 'Experiment' tab.

6. MAINTENANCE

This section covers the care of your Dualo 32 R2, including cleaning, environmental conditions, and disassembly.

Cleaning

Day-to-Day

Wipe the external surface with a damp, soft, lint-free cloth. Dry with another soft, lint-free cloth.

Notes: Avoid abrasive cloths, towels, or paper towels. Before cleaning, unplug all external power sources, devices, and cables. Prevent moisture from entering any openings.

Deep Cleaning Wells

Automated background subtraction minimizes the impact of low-level fluorescence contamination. For wells with high levels of fluorescent substances, contact technical support for guidance.

Environmental Conditions

Your Work Space

Place the Dualo 32 R2 on a flat, dry surface away from drafts. Do not install it in direct airflow from air conditioners or fans, or in dusty environments. Ensure airflow around the instrument, including the heated lid and vents, is not obstructed.

Environmental Operating Conditions

Environmental Storing / Transporting / Packing Conditions

Disassembly

The Dualo 32 R2 contains no user-serviceable components. Any disassembly will void all warranties.

7. TROUBLESHOOTING

This section helps you troubleshoot issues with your Dualo 32 R2.

Consumables

• 8 tube-strips, Product No. Z 100 87
• 8-Cap strip, Product No. Z 100 89

FAQ

Can I leave my Dualo 32 R2 powered on overnight?

Yes. It is safe to leave your Dualo 32 R2 powered on overnight; it will enter a low power standby mode.

Where is the threshold for determining Cq values in Auto Quant?

Modern Cq value determination is model-based, not threshold-based. Auto Quant fits a PCR amplification model to the observed fluorescence data, enabling estimation of parameters including Cq values.

Error Messages

Errors are displayed as messages in the Status Bar of the 32 R2 software or by the instrument's display LEDs.

If an error occurs, ensure the instrument is in a suitable lab environment, tubes are loaded correctly, the lid is fitted properly, and all cables are securely attached. If the error persists, contact technical support.

Error Messages - Displayed in Status Bar

Error Log Files

In the event of a problem, the software can generate log files to aid diagnosis.

Software Errors

For software issues, go to Configuration > About and select 'Save Software Logs to File'.

Hardware Errors

For hardware faults, go to Configuration > Dualo 32 R2 and select 'Retrieve Instrument Report Files'. Send the error file, serial number, and the experiment file containing the error to Technical Support. Experiments that did not complete or were aborted can also be sent.

WARRANTY AND DISCLAIMER OF LIABILITY

Limited Warranty and Disclaimer of Liability: Hygiena® Diagnostics GmbH warrants this product against defects in materials and workmanship for 12 months, provided the following conditions are met:

1. The product is used according to the guidelines and instructions in the product literature.
2. Hygiena Diagnostics GmbH does not warrant against defects resulting from materials or workmanship not provided by Hygiena Diagnostics GmbH, misuse, use contrary to instructions, or contamination from improper storage or handling.
3. Warranties of merchantability and fitness for a particular purpose (written, oral, expressed, or implied) are limited to one year from the date of manufacture. No other warranties extend beyond those described.
4. Hygiena Diagnostics GmbH is not responsible for undertakings, representations, or warranties made by dealers or distributors unless expressed in writing by an officer of Hygiena Diagnostics GmbH.
5. Hygiena Diagnostics GmbH assumes no responsibility for incidental or consequential damages, including but not limited to loss of use, labor costs for removal/replacement, loss of time, inconvenience, expenses for calls, shipping, property damage, loss of revenue, personal injuries, or wrongful death.
6. Hygiena Diagnostics GmbH reserves the right to replace or credit any returned modules under this warranty.

SUPPLEMENTARY INFORMATION

Ordering Information

Contact your local Hygiena Diagnostics GmbH representative for ordering information.

Trademarks

Dualo 32® is a trademark of Hygiena Diagnostics GmbH. Hygiena® is a registered trademark of Hygiena. Other brand or product names are trademarks of their respective holders.

Change Index

• Version 1, June 2020: First version of the package insert
• Revision A, July 2023: Rebranding and numbering

Contact Information

Manufactured in the United Kingdom for Hygiena Diagnostics GmbH

Hermannswerder 17
14473 Potsdam
Germany

Website: www.hygiena.com

For inquiries, please contact your nearest Hygiena office: www.hygiena.com/Support