What is the difference between continuous melt curves and step and hold melt curves on the Applied Biosystems Real-Time PCR Instruments?
Continuous melt curve: increases the temperature by the ramp increment (degrees C/sec) so that the temperature will change at a constant rate set in the protocol.
Step and Hold melt curve: increases the temperature by the ramp increment (degrees C) and then holds at that temperature for the time specified by the user.
The amount of data collected will depend on the filter(s) selected. The more filters you use, the longer the acquisition will take and the fewer data points will be collected.
I am unable to download the current software update for the QuantStudio 5 Real-Time PCR System from the Thermo Fisher website. Why?
We recommend trying different browsers (Google Chrome, Firefox, and Microsoft Edge) and/or removing cookies from your browser. If this does not resolve the issue, please contact Technical Support at techsupport@thermofisher.com.
What is the admin pin for QuantStudio 3 and 5 Real-Time PCR Systems?
If the admin PIN has not been changed, 1234 should work. If it does not work, you can also try 0000.
However, if the admin PIN has been changed and none of the above PINs work, follow the instructions in the QuantStudio 3 and 5 Real-Time PCR Systems manual, page 100.
Can I import a standard curve for my real-time PCR data into the Design & Analysis Software (v2.x)?
It is possible to import a standard curve into the Design & Analysis Software (v2.x) if you are analyzing results from one of the following Applied Biosystems real-time PCR instruments:
• 7900HT Fast Real-Time PCR System
• 7500/7500 Fast Real-Time PCR System
• StepOne/StepOne Plus Real-Time PCR System
• ViiA 7 Real Time PCR System
• QuantStudio 1 Real-Time PCR System
• QuantStudio 3 & 5 Real-Time PCR System
• QuantStudio 6 & 7 Flex Real-Time PCR System
• QuantStudio 6 & 7 Pro Real-Time PCR Systems
• QuantStudio 12K Flex Real-Time PCR system
Follow the instructions below to import a standard curve for analysis in the Design & Analysis Software (v2.x):
1. Open the data file in the Design & Analysis Software (v2.x).
2. Click on Actions.
3. Click on "Standard Curve Analysis Setting".
4. Select "External Standard Curves" and click on "Import".
5. Browse to the proper .csv file and click "Open".
6. Click "Apply".
Can I import a standard curve for my real-time PCR data into the QuantStudio Design & Analysis Software (v1.x)?
If you are analyzing results from the QuantStudio 1 Real-Time PCR System, QuantStudio 3 Real-Time PCR System, or QuantStudio 5 Real-Time PCR System, it is possible to import a standard curve into the QuantStudio Design & Analysis Software (v1.x) following the instructions below:
1. Open the data file in the QuantStudio Design & Analysis Software (v1.x).
2. Go to Analysis Settings.
3. Click on the "Standard Curve Settings" tab.
4. Select the option to "Use a standard curve imported from another experiment" and click on "Import".
5. Browse to and highlight the proper data file and click "Select".
6. Click Apply.
Can you provide some tips for setting up an absolute quantification/standard curve on the QuantStudio 3 or 5 Real-Time PCR instrument?
Please review this guided tutorial that explains how to set up an Absolute Quantification/Standard Curve experiment run on your QuantStudio 3 or 5 Real-Time PCR instrument: https://www.brainshark.com/thermofisher/QS35AQSetupTutorial
How do I set up the QuantStudio 3 or QuantStudio 5 Real-Time PCR System for WiFi access?
To set up the QuantStudio 3 or QuantStudio 5 Real-Time PCR System for WiFi capability, we recommend following the instructions in the manual (https://tools.thermofisher.com/content/sfs/manuals/MAN0010407_QuantStudio3_5_InstallUseMaint_UG.pdf). You will also need to purchase the High Power USB WiFi Module (Cat. No. A26774).
Can I use templates from other instruments to start a run in the QuantStudio 3 or QuantStudio 5 Real-Time PCR System?
Yes, you can use templates from other compatible systems, if you use the Thermo Fisher Cloud Design and Analysis module. Use the function “New Experiment --> From File” to import the .edt file.
I am having trouble connecting to my Thermo Fisher Cloud account, what could be wrong?
Check if the Cloud Monitor is on. Go to Settings, then Maintenance and Service, then Monitoring. Turn CloudSuite Monitor ‘ON'. This needs to be set to ‘ON', otherwise the instrument will not look out to connect to your account even if you are logged in.
Are my run files from the QuantStudio 3 or 5 Real-Time PCR System compatible with TaqMan Genotyper Software?
No. You can use the QuantStudio Design and Analysis Desktop Software, or for multi-plate analysis of genotyping data, please use the Applied Biosystems qPCR Analysis Module (GT application) with your Thermo Fisher Cloud account.
Are my run files from the QuantStudio 3 or 5 Real-Time PCR System compatible with ExpressionSuite Software?
Yes, ExpressionSuite Software v1.1 is compatible with files from the QuantStudio 3 and 5 Real-Time PCR Systems..
I have a qPCR master mix from another vendor that has ROX passive reference dye in a separate tube. How much should I add for my reactions in the QuantStudio 3 and 5 Real Time PCR Systems?
If you are using a master mix that has ROX provided in a separate tube, use a low ROX concentration (50 nM, final well volume).
How do I analyze multiple ddCt files together from a QuantStudio 3 or 5 Real-Time PCR System?
Use the Applied Biosystems qPCR Analysis Module (RQ application) with your Thermo Fisher Cloud account.
How do I start a qPCR run on the QuantStudio 3 or 5 Real-Time PCR System using the cloud?
1. Set up the run in the cloud from the Design and Analysis application within the Applied Biosystems qPCR Analysis Modules.
2. Save the experiment before starting a run: The file name was created from the experiment Name entered in the Properties tab. However, changes to the experiment Name in the Properties tab after the first save do not update the file name. To change the file name before a run, select Actions, then Save As.
3. Go to an available cloud-connected instrument, enter your PIN to access this file, and start the run by choosing ‘Open Template'. Following a run, view experiment details, run duration, and instrument and software information in the Run Control screen of the Run tab.
Can I start a run on the QuantStudio 3 or 5 Real-Time PCR System from the Thermo Fisher Cloud Design and Analysis application?
You can set up your run in the Design and Analysis application, but to actually start the run, you will have to go to the instrument touchscreen.
Can I monitor my qPCR run from the QuantStudio 3 or 5 Real-Time PCR System in real time?
Yes, you have several options for this. You can monitor the run in real time from:
- The instrument touchscreen
- The QuantStudio Design and Analysis Desktop Software
- Online using the Design and Analysis application within Thermo Fisher Connect
- Online using the Instrument Connect App on your Android or Apple mobile device - for more on this, please see here (https://www.thermofisher.com/us/en/home/life-science/pcr/real-time-pcr/real-time-pcr-instruments/quantstudio-3-5-real-time-pcr-system/connectivity-quantstudio-3-5-qpcr-data-analysis-software.html).
With the QuantStudio 3 or QuantStudio 5 Real-Time PCR System, I want my file to go to my Thermo Fisher Cloud account once the run is complete for analysis. Is there a way for it to automatically upload?
Yes. To have the run file automatically upload to your Thermo Fisher Cloud account once complete, you will need to set the file destination on the touchscreen, prior to starting the run:
1. Before you start the run, select the ‘Properties' tab from an open experiment
2. Touch ‘Edit', Data Destination - Choose USB or Cloud
Alternatively, you could use Cloud Connect Utility to have all files from a specific hard drive folder location automatically upload to the cloud (for use with the QuantStudio Design and Analysis Desktop Software).
How do I open the drawer in the QuantStudio 3 or QuantStudio 5 Real-Time PCR System?
The block drawer is opened and closed by using the ‘eject' button on the touchscreen. Do not push or pull on the instrument drawer.
When do I have to calibrate the QuantStudio 3 and QuantStudio 5 Real Time PCR Systems?
When your new QuantStudio 3 or QuantStudio 5 Real-Time PCR System arrives in your lab, it will be factory calibrated. The recommended calibration schedule is to recalibrate every 2 years.
Can I purchase or use the QuantStudio 3 or QuantStudio 5 Real Time PCR System without a computer?
Yes, you can purchase the QuantStudio 3 or QuantStudio 5 Real Time PCR System without a computer. In this configuration, you can run the instrument through the touchscreen or the Design and Analysis application from Thermo Fisher Cloud.
Do the QuantStudio 3 and QuantStudio 5 Real-Time PCR Systems have the ability for security and auditing software?
Yes, the QuantStudio 5 Real-Time PCR System has SAE (Security/Auditing/e-Signature) features that can be enabled in the QuantStudio Design and Analysis Desktop Software.
What is the difference between the Applied Biosystems QuantStudio 3 and Applied Biosystems QuantStudio 5 Real-Time PCR Systems?
The QuantStudio 3 Real-Time PCR System is a 4-color instrument with a 3-zone VeriFlex block, while the QuantStudio 5 Real-Time PCR System is a 6-color instrument with 6-VeriFlex zones in a 96-well block, (or 5-color format, no VeriFlex with 384-well block).
