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Optimize your experiments to get the best results. We’ve compiled a detailed knowledgebase of the top tips and tricks to meet your research needs.
View the relevant questions below:
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Recommended Maintenance Schedule | |
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Power on/off the computer controlling the instrument | Weekly |
Check computer disk space. If necessary, archive or back up your experiment files and instrument settings | Weekly |
Background Calibration | Every month |
Run disk cleanup and disk defragmentation | Every month |
Perform an instrument self test | Every month |
Pure dye calibrations | Every 12 months |
ROI calibration | Every 12 months |
Uniformity calibration | Every 12 months |
Normalization calibration | Every 12 months |
RNase P instrument verification | After the instrument has been moved, or as needed to verify instrument performance |
The uniformity calibration generates data that allows the software to compensate for the physical effects of the QuantStudio™ 12K Flex Real-Time PCR System filters using the ROI plate. The normalization calibration generates factors that the software uses when comparing data from multiple QuantStudio™ instruments within a study. There are 2 normalization plates provided in the calibration kit, one containing FAM™/ROX™ dye and the other with VIC®/ROX™ dye.
All the calibration plates can be stored and reused three times for up to 6 months after you first open them, so make sure to return them to their original packaging and return them to –20°C storage until the next use. (Note: Due to the small volumes, calibration array cards cannot be reused. Please discard them after use.) If needed, you can make your own background plate using deionized water. Please follow the directions in the QuantStudio™ 12K Flex Real-Time PCR System Maintenance and Administration Guide (Appendix C) for more details.
Please ask your dye manufacturer for recommendations. If there is no specific recommendation available, try TE Buffer (pH 8.0) to start.
The RNaseP verification plate contains template, master mix, and a TaqMan® assay for RNaseP. It is used to verify that the instrument is performing to specifications. If you have reason to suspect there is something wrong with the instrument, or if you want to rule out a chemistry issue, the RNaseP plate is a good way to test the system. The RNaseP verification plate is a single-use plate.
We recommend performing the calibration for the OpenArray™ block every 12 months. This can be done using the QuantStudio™ 12K Flex OpenArray™ Calibration Kit (Cat. No. 4478601).
Instrument Specifications | |
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Block Options (Interchangeable) | 96-well (standard), 96-well (Fast), 384-well, TaqMan® Array Card, OpenArray® plate (4 plates) |
Sensitivity | Down to 1 copy |
Dynamic Range | 9 logs of linear dynamic range |
Calibrated Dyes | FAM™, SYBR®, VIC®, ROX™, NED™, TAMRA™ dyes |
Detection Method | SYBR®, primer-probe detection |
Resolution | Detect changes as little as 1.5-fold |
Reaction Volume Range | 10–100 µL (96-well standard) 15–30 µL (96-well Fast) 5–20 µL (384-well) ~1 µL (TaqMan® Array Card) 33 nL (OpenArray®Plate) |
Reaction Speed | Fast or standard |
Optics | 6 excitation filters (450–670 nm), OptiFlex™ system (enhanced fluorescence detection), 6 emission filters (500–720 nm) |
Temperature Range | 4–99.9°C |
Run Time | 2 hr (standard mode) ~35 min (Fast mode) |
Regulatory Statement | For Research Use Only. Not for use in diagnostic procedures. |
Temperature Accuracy | ±0.25°C (between 35°C and 95°C, after 3 min) |
Temperature Uniformity | ±0.5°C (after 30 sec) |
Thermal Cycling System | Peltier-based system |
Available Applications | Gene expression, genotyping, copy number variation, HRM, protein thermal shift, protein detection, mutation detection, miRNA, presence/absence, digital PCR |
Dimensions | 50.5 cm (W) x 67.2 cm (D) x 73.8 cm (H) |
Weight | 69.5 kg (152 lb) |
Remote Monitoring | Yes (up to 10 instruments) |
On-Board Memory | Yes (up to 100 experiments) |
Setup Configurations | PC-controlled or run directly from touch screen |
The QuantStudio™ 12K Flex Real-Time PCR System features a 6-color filter set that supports all Life Technologies™ dyes. Custom dyes that excite between 455 and 672 nm and read between 505 and 723 nm can also be used, although you will have to calibrate the system first for any new dye.
In a touchdown PCR experiment, you will change either the temperature or the time of a particular PCR step with every cycle. Most commonly, the annealing temperature is adjusted throughout the experiment, such that the specificity is higher in the early cycles and the efficiency in the later cycles.
In this example, we will set the method to do the following:
40 cycles |
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You will need an oligo sequence with the custom dye but without the quencher molecule. For each custom dye, prepare samples in a concentration range of 100–2,000 nM, choosing a 2–3-fold difference in dilution points. Use either the background buffer provided in the calibration kit or your own buffer to dilute the oligo. A full plate is not needed; see the example below.
In this example, sample concentrations would be 100, 200, 400, 800, and 1,600 (20 µL/well).
Note: The volume would be the same for a 96-well or 384-well plate.
The QuantStudio™ 12K Flex Real-Time PCR System can store up to 100 runs on the instrument itself.
The following volumes are supported for each instrument block:
Yes. The QuantStudio™ 12K Flex Real-Time PCR System can collect data at multiple steps in the amplification stage. You will need to turn on the data collection for every step of interest. In the analysis, only one set of data can be displayed at a time. To change the data set used for analysis, go to Analysis Settings → Ct Settings and change the dropdown under ‘Data Step Selection’.
Run files will be saved to both the instrument and the connected computer. On the connected computer, files will be saved to the default data folder, unless you change it. To find or change the default folder, go to Tools → Preferences → Defaults. Here you will see a Data Folder and an Import Folder. The default location is shown. If you want files to be saved to (or open from) a different location, click ‘Browse’ and choose the new folder.
Follow the directions in this blog post to transfer your license to another computer.
To get log files from the QuantStudio™ 12K Flex Real-Time PCR instrument,
To get log files from the QuantStudio™ 12K Flex Real-Time PCR software on the client computer,
MicroAmp 96-Well Tray for VeriFlex Blocks (Cat. No. 4379983) is needed to load 1 or 2 MicroAmp Optical 8-Tube Strips with Attached Optical Caps on the ProFlex, SimpliAmp, MiniAmp, MiniAmp Plus, and Veriti Thermal Cycler 96-well blocks, and QuantStudio 3 and QuantStudio 5 blocks.
The MicroAmp Optical 8-Tube Strip with Attached Optical Caps is compatible with the following instruments:
- ProFlex 3 x 32-well PCR System
- ProFlex 96-well PCR System
- Veriti 96-well Thermal Cycler
- SimpliAmp Thermal Cycler
- MiniAmp Thermal Cycler
- MiniAmp Plus Thermal Cycler
- 2720 Thermal Cycler
- GeneAmp PCR System 9700, 96-well
- 7000 PCR System
- 7300 Real-Time PCR System
- 7500 Real-Time PCR system
- QuantStudio Real-Time PCR Systems
- ViiA7 Real-Time PCR System, 96-well
The MicroAmp 8-Tube Strip with Attached Domed Caps is compatible with the following instruments:
- ProFlex 3 x 32-well PCR System
- ProFlex 96-well
- Veriti 96-well Thermal Cycler
- SimpliAmp Thermal Cycler
- MiniAmp Thermal Cycler
- MiniAmp Plus Thermal Cycler
- 2720 Thermal Cycler
- GeneAmp PCR System 9700, 96-well
MicroAmp 96-Well Tray for VeriFlex Blocks (Cat. No. 4379983) is needed to load 1 or 2 MicroAmp Optical 8-Tube Strips with Attached Domed Caps on the ProFlex, SimpliAmp, MiniAmp, MiniAmp Plus, and Veriti Thermal Cycler 96-well blocks.
Yes. The tubes are DNase/RNase/Human DNA free, as they are manufactured in a Class 100K ISO certified clean room UK production facility.
Yes. The tubes are DNase/RNase/Human DNA free, as they are manufactured in a Class 100K ISO certified clean room UK production facility.
These new strips have new features including a graduated 20 μL measuring mark for visual verification and dual end tabs for better labeling and handling. In addition, the individual caps help prevent cross contamination and reduce sample evaporation.
These new strips have new features including a graduated 20 μL measuring mark for visual verification and dual end tabs for better labeling and handling. In addition, the individual caps help prevent cross contamination and reduce sample evaporation.
Please refer to the table on Page 2 of this flyer for compatibility information.
Please refer to the table on Page 2 of this flyer for compatibility information.
Yes. The tubes are DNase/RNase/Human DNA free, as they are manufactured in a Class 100K ISO certified clean room UK production facility.
Yes. The tubes are DNase/RNase/Human DNA free, as they are manufactured in a Class 100K ISO certified clean room UK production facility.
These new strips have new features including a graduated 20 μL measuring mark for visual verification and dual end tabs for better labeling and handling. In addition, the individual caps help prevent cross contamination and reduce sample evaporation.
These new strips have new features including a graduated 20 μL measuring mark for visual verification and dual end tabs for better labeling and handling. In addition, the individual caps help prevent cross contamination and reduce sample evaporation.
Additional OpenArray® plates can be run off-line using a flat-block thermal cycler. The two thermal cyclers we recommend are the GeneAmp® PCR System 9700 and the ProFlex™ PCR System.
You need an OpenArray® Plate Adaptor, Cat. No. 4472306. You do not need a compression pad. This part number works with both the GeneAmp® PCR System 9700 and the ProFlex™ PCR System.
You need an OpenArray® Plate Adaptor, Cat. No. 4426824, and a compression pad, Cat. No. 442825.
For more detailed instructions, refer to the Dual Flat Block GeneAmp® PCR System 9700 User Guide.
These instructions may be found on page 5 of the Dual Flat Block GeneAmp® PCR System 9700 User Guide.
93°C for 10 min, ramp rate 100%
50 cycles of:
95°C for 45 sec, 84% ramp rate
94°C for 13 sec, 100% ramp rate
53°C for 2 min 14 sec, 44% ramp rate
25°C for 2 min, 100% ramp rate
Optional: Add a post-PCR overnight hold as shown below:
4°C for 99 min 59 sec, 100% ramp rate
These instructions may be found on page 79 of the ProFlex™ PCR System User Guide.
Reaction volume: 33 nL
Cover temperature: 70°C
94°C for 600 sec, ramp rate 1.3
50 cycles of:
99°C for 30 seconds, ramp rate 1.3
54°C for 21 seconds, ramp rate 1.1
57°C for 90 seconds, ramp rate 0.1
To perform pre-read and post-read, choose “Get Plate IDs” and input the setup file either with Genotyping.edt or .spf for your runs.
Check the box for “Include Pre-read” and uncheck “Include Amplification”.
Edit the file location and the file name and start the run.
After your pre-read run has ended, you can either keep the run console on to continue with the post-read, or you can close it and perform other pre-read runs.
If you closed the run console and wish to perform the post-read, put the OpenArray in the same position as it was when you did the pre-read and choose “Get Plate IDs” again. Select the .eds file that you had performed pre-read as the setup file and start the run. Note: it is important here that you select the .eds file, NOT the .spf or .edt.
If you have placed the array at the wrong position, the software will prompt you to select the correct position.
For genotyping data, we recommend to use TaqMan® Genotyper™ software. For relative Quantitation, we recommend to use ExpressionSuite software.
The ‘reveal traces’ feature for genotyping runs allows you to trace the clusters throughout the PCR process. For some assays you may find better cluster discrimination at, say, cycle 40, as opposed to cycle 50. To use this feature, the amplification data need to have been collected for the run.
The plot on the left shows a genotyping data set at a full 40 cycles. The plot on the right is the same data set after revealing traces (grey lines). The clusters can be traced back by moving the cycle bar. Notice that one point is no longer called at the earlier cycle set point.
You can download your SPF plate files from here.
For Research Use Only. Not for use in diagnostic procedures.