QuantStudio™ Absolute Q™ Digital PCR System, desktop - FAQs

View additional product information for QuantStudio™ Absolute Q™ Digital PCR System, desktop - FAQs (A52864)

24 product FAQs found

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.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

What is the limit of detection for the QuantStudio Absolute Q Digital PCR System (Cat. No. A52864)?

The QuantStudio Absolute Q Digital PCR Instrument (Cat. No. A52864) uses QuantStudio Q MAP16 Digital PCR Plates that contain 16 digital PCR microreaction chamber arrays, each containing 20,480 fixed volume microreaction chambers of 432 ρL (0.000432 µL). If there is 1 copy of gene target in the tested sample, and the sample is using 1 array of the MAP16 plate, then the theoretical lower limit of detection (lowLOD) for the Absolute Q can be approximated by the following calculation:

lowLOD for Absolute Q = (1 copy)/ (0.000432 µL X 20,480 chambers) = 0.113 copies per µL

If a sample is run in triplicate, and there are 3 positive chambers, the lowLOD in practice for Absolute Q can be calculated using the following calculation:

lowLOD for Absolute Q = (3 copies)/ (0.000432 µL X 20,480 chambers) = 0.339 copies per µL

The QuantStudio Absolute Q has the sensitivity of detecting the presence of a single copy of target sequence within the 9 µL reaction volume, which would result in a detected concentration of ~0.1 copies per µL. This detection limit can be reduced even further by using the software’s digital pooling feature to increase the total analyzed volume. The ability of the instrument to detect at this level will be impacted by various components including the assay efficiency and reagents used in the reaction. The LOD is also dependent on a given assay’s specificity as well as the potential background fluorescence signal from the sample matrix, which may affect the LOD achieved in practice. The Absolute Q software’s unique false positive detection capability can enhance an assay’s performance, especially when low target concentrations are investigated.

It is important to note that the LOD should be determined empirically by the user for their specific experimental conditions.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Applications Support Center.

What optical dyes does QuantStudio Absolute Q Digital PCR System (Cat. No. A52864) support?

QuantStudio Absolute Q Digital PCR System (Cat. No. A52864) supports the following dyes: FAM™ dye, VIC™ dye, HEX™ dye, ABY™ dye, ROX™ dye, CY™5 dye, and JUN™ dye.

Please refer to page 13 of the QuantStudio Absolute Q Digital PCR System User Guide for Installation, Use and Maintenance for more information on supported optical dyes and channel selection.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

I finished running my experiment on the QuantStudio Absolute Q Digital PCR instrument but just realized that I had selected the wrong dyes during setup. Can I change the dyes that were used after the run?

If dye channels are turned off before the run, then data will not be collected for them as the run progresses. This means that changing the dyes after the run will not be possible.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

I ran my experiment using the QuantStudio Absolute Q Digital PCR System but did not actually use dyes in all the channels. Is there a way to only see those dye channels that were used?

Unused dye channels can be turned off after the run by going to the SETUP tab and clicking EDIT SETUP and then EDIT GROUPS. Here, the Analysis can be changed to “Not Used.” After clicking SAVE twice, the data will be reanalyzed and the unused dye channels will not be present in the analysis.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

I'm using the QuantStudio Absolute Q Digital PCR System and the software is saying that the concentration is NaN. What does that mean?

NaN stands for “not a number”. The software will display NaN instead of an actual number when it has detected some issue during the analysis of the array images.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

With the QuantStudio Absolute Q Digital PCR instrument, the run data didn't automatically transfer after the run completed. How can I get the data?

We would recommend restarting the software. If the run still doesn't transfer, we would recommend closing the software, rebooting the instrument, and then starting the software. If further assistance is needed, please contact Technical Support at techsupport@thermofisher.com.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

With the QuantStudio Absolute Q Digital PCR System, how much sample can be loaded into each well?

9 µL of sample is loaded into each well along with 15 µL of isolation buffer. Loading more or less of either the sample or isolation buffer will affect filling of the microchambers and the run results.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

How do I know how much sample to use with the QuantStudio Absolute Q Digital PCR System?

Please see the sections on determining the optimal dilution in Chapter 2 of the QuantStudio Absolute Q Digital PCR System Installation, Use, and Maintenance Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0025621_QS_Absolute_Q_Digital_PCR_System_UG_RUO_EN.pdf).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

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How many samples can be run on the QuantStudio Absolute Q Digital PCR instrument at one time?

The QuantStudio Absolute Q MAP16 Digital PCR Plate can accommodate up to 16 individual samples. Loading the same sample in multiple microarrays as replicates or for the purpose of pooling results will decrease the number of individual samples that can be run on the same plate.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

How many QuantStudio Absolute Q Digital PCR instruments can be managed at the same time using the QuantStudio Absolute Q Digital PCR Software?

QuantStudio Absolute Q Digital PCR Software can only manage one QuantStudio Absolute Q Digital PCR instrument at a time.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

How long does a run on the QuantStudio Absolute Q Digital PCR System take?

A result can be provided in less than 90 mins depending on the protocol that is used.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Are there any special precautions when loading the QuantStudio Absolute Q MAP16 Digital PCR Plate?

Yes. Here are some precautions to take:
- Do not allow the pipette tip to come in contact with the bottom of the well or puncture the thin membrane that resides at the bottom of the well.
- Be careful not to induce bubbles when pipetting into the wells.
- Do not mix the isolation buffer and sample.
- Ensure that the gaskets are oriented the correct way and are pushed all the way on.

For more detailed instructions, please see the guidelines in Chapter 2 of the QuantStudio Absolute Q Digital PCR System Installation, Use, and Maintenance Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0025621_QS_Absolute_Q_Digital_PCR_System_UG_RUO_EN.pdf).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

What master mix can I use with the QuantStudio Absolute Q Digital PCR System?

The QuantStudio Absolute Q Digital PCR System is meant to be used with the Absolute Q DNA Digital PCR Master Mix (5X) (Cat. No. A52490), Absolute Q 1-step RT-dPCR Master Mix (4X) (Cat. No. A55146), or Absolute Q Universal DNA Digital PCR Master Mix (5X) (Cat. No. A72710). Other master mixes have not been validated with this system.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

How do I find the .zst file for the QuantStudio Absolute Q Digital PCR System?

The .zst data files are not automatically generated. To create a .zst file, select the run by checking the box to the left of the run name and then click on the “Export Run” button that appears. You will be asked if that is the run you would like to export and then would need to select a location to export the file to.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

What dyes are supported by the QuantStudio Absolute Q Digital PCR System?

The QuantStudio Absolute Q Digital PCR System has filters that support FAM, HEX (not field calibrated), VIC, ABY, ROX (QC channel), CY5, and JUN dyes.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Does the QuantStudio Absolute Q instrument use channels (e.g., ROX dye) to determine which compartments have a reaction?

Yes, ROX dye is the passive reference dye for QC (Quality Control).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Do I need to use the entire QuantStudio Absolute Q MAP16 Digital PCR Plate at one time?

No. Microarrays will be used in columns of 4 and unused columns can be used at a future time.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

What does the QuantStudio Absolute Q MAP16 Digital PCR Plate look like?

The QuantStudio Absolute Q MAP16 Digital PCR Plate is a microinjection-molded plate which is 85.5 mm x 128 mm and consists of 16 microarrays. Each microarray consists of approximately 20,000 microchambers, each accommodating 432 pL (picoliter) volume of reaction. See image on page 29 of the QuantStudio Absolute Q Digital PCR System Installation, Use, and Maintenance Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0025621_QS_Absolute_Q_Digital_PCR_System_UG_RUO_EN.pdf).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

What are the electrical and environmental specifications for the QuantStudio Absolute Q Digital PCR System?

For QuantStudio Absolute Q instrument specifications, please see Appendix F of the QuantStudio Absolute Q Digital PCR System Installation, Use, and Maintenance Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0025621_QS_Absolute_Q_Digital_PCR_System_UG_RUO_EN.pdf).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

How many QuantStudio Absolute Q instruments can be managed through QuantStudio Absolute Q Software?

QuantStudio Absolute Q Software van only manage one QuantStudio Absolute Q instrument at a time.

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

Does the QuantStudio Absolute Q instrument use channels (e.g., ROX) to determine which compartments have a reaction?

Yes, ROX is the passive reference dye for QC (Quality Control).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

For the QuantStudio Absolute Q Digital PCR System, what does MAP mean or stand for?

MAP stands for patented Microfluidic Array Plate technology. The QuantStudio Absolute Q Digital PCR instrument uses QuantStudio Absolute Q MAP16 Digital PCR Plates (MAP plates) for loading samples and running experiments. The MAP plates use microinjection-molded plate technology to overcome common reagent distribution challenges of inconsistency and high dead volume. This novel approach provides many benefits, including precise micro-chamber volume, consistent sample digitization, and greater than 95% of the loaded volume analyzed:
- 16 samples per plate
- 20,000 micro-chambers per reaction
- <5% dead volume

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.

For the QuantStudio Absolute Q Digital PCR System, what is the volume per compartment?

Each microchamber holds 432 picoliters (pL).

Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.