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查看更多产品信息 QuantStudio™ 1 Real-Time PCR System, 96-well, 0.2 mL, laptop - FAQs (A40427)
17 个常见问题解答
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.
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".
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.
It is possible that the instrument is not connected to the computer and is not linked with the Thermo Fisher Cloud. Please see page 38 of the QuantStudio 1 Real-Time PCR System User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017853_QS1%20UG.pdf) for instructions to link an instrument profile with the Thermo Fisher Cloud. Although default installation instructions recommend using an Ethernet cable, it can also be done wirelessly using a High Power USB WiFi Module, Cat. No. A26774 (https://www.thermofisher.com/order/catalog/product/A26774).
If you experience connection issues between the instrument and computer, it might be useful setting a static IP address on both the instrument and computer. On the computer, you can use online help, such as this one (https://kb.netgear.com/27476/How-do-I-set-a-static-IP-address-in-Windows) to set the static IP address. Here are the details for the settings:
- Computer IP: 10.0.0.2
- Instrument IP: 10.0.0.3
- Subnet: 255.255.255.0
- DNS1: 8.8.8.8
- DNS2: 8.8.4.4
- Default gateway: 10.0.0.1
On the instrument, the IP address can be found under Settings and About Instrument. If you see an IP address but are still having network issues, try to PING the IP Address in a command window of the computer:
- Select the windows start button and select the RUN command.
- Open the Run command and type in CMD.
- This will open the Command window.
- Type the word "PING", space, followed by the IP address and press enter.
- You should see 4 packets of data sent, 4 received, and 0 lost.
If you PING the IP Address successfully (4 sent, 4 received, 0 lost), but still cannot connect, it means that one of the needed ports is still not open. Please contact your IT department and review the instrument checklist.
If you PING the IP address unsuccessfully, please send us the serial number of the instrument so that we can contact your local engineer.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
In an effort to reduce environmental impact, we no longer supply the software on the CD. Please download it from here (https://www.thermofisher.com/uk/en/home/global/forms/life-science/quantstudio-software-download.html).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
We recommend storing no more than 1000 run files in the instrument. Please transfer EDS files from the instrument home screen, in the Settings > Run History, as shown on page 48 of the QuantStudio 1 Real-Time PCR System User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017853_QS1%20UG.pdf). Only Administrators can delete all files, however.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Although we do not offer Security and Audit module (SAE) to assist with the 21 CFR p11 compliance for the QuantStudio 1 instrument, it can be used with guest, user, and administrator accounts:
- Guests (users not signed in) can only view guest run records.
- Users signed in with password into their instrument profiles can also view their own run records.
- Administrators can view all run record.
In addition, the touchscreen can be locked and protected by PIN code (please go to My Profile, Lock Screen). The templates can also be locked by password for editing, which will lead to generation of locked run files.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
To see the red threshold line in the amplification plot, please go to analysis settings and select Baseline Threshold instead of Relative Threshold. Also, please select a single target in the amplification plot, from the drop-down menu.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Calibrating the instrument specifically for the dye being used may improve assay efficiency. The HEX dye needs to be custom calibrated on the QuantStudio 1 instrument. Please see page 62 of the QuantStudio 1 Real-Time PCR System User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017853_QS1%20UG.pdf) for further instructions. You may also refer to our Custom Dye Calibration of Applied Biosystems Real-Time PCR Instruments User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/CustomDyeCalibration_AB_RealTimePCR_UB.pdf).
For the custom dye calibration, we recommend using either a pure HEX dye purchased from the vendor producing the HEX probe or a single-labeled HEX oligo. Custom dye calibration using a labeled HEX oligo is easier to manipulate and dilute and follows the same procedure as if using the pure dye. The length or sequence of the oligo is not important and desalted purification method would suffice. No other components are required. The HEX oligo can be ordered as a 5' Fluorescent Labeled Single Primer (https://www.thermofisher.com/order/custom-oligo/fluorescent-labeled-primers) or Custom Standard DNA Oligo (https://www.thermofisher.com/order/custom-standard-oligo).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
The internet connection would allow monitoring the run via a mobile app, uploading templates from the Thermo Fisher Cloud on any other computer, and getting the most recent updates of firmware and software, etc. However, internet connection is not necessary to start the run and perform the analysis.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
The QuantStudio 1 instrument has filters to perform a duplex reaction with FAM/VIC and ROX as the passive reference dye. Alternatively, a triplex reaction can be carried out using FAM/VIC/JUN without a passive reference dye. Please see the figure on page 10 of the QuantStudio 1 Real-Time PCR System User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017853_QS1%20UG.pdf).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
The QuantStudio 1 instrument has a "pause" option for holding the cycling. This pause can be actually scheduled in the middle of the run. Please see page 46 of the QuantStudio 1 Real-Time PCR System User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017853_QS1%20UG.pdf) for further details. It is also possible to pause a run, set a pause temperature, and then open the drawer to access the plate during the run using "unlock drawer" option.
Please note that the plate temperature can reach 100 degrees C during the run. If you want to access the plate during a run pause, enter "room temperature" as the pause temperature and allow the plate to cool to room temperature before handling.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Yes, it is possible. Please use local area network (LAN) connection and see page 17 of the QuantStudio 1 Real-Time PCR System User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0017853_QS1%20UG.pdf) for supported configuration options. However, it is not possible to connect more than one instrument directly to one computer (supplied by Thermo Fisher Scientific) using wired connection due to single Ethernet port availability on the computer supplied by us. Also, please note that starting a run simultaneously from the same computer on two connected instruments is not yet fully validated.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Yes, it is possible. Please use the following settings to start the run: Melt Curve experiment type, ROX filter, and NFQ-MGB probe type and recommendations as described in the Protein Thermal Shift User Guide (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/cms_095141.pdf).
The Protein Thermal Shift Software v1.4 (https://www.thermofisher.com/de/en/home/technical-resources/software-downloads/protein-thermal-shift-software.html) is now compatible with the QuantStudio 1 instrument and Windows 10.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Yes, it is possible. Please calibrate the instrument for MeltDoctor dye using MeltDoctor HRM Calibration Standard (20X), Cat. No. 4425562 (https://www.thermofisher.com/order/catalog/product/4425562) and the custom dye calibration protocol. The run should be performed with standard melt curve experiment settings. For data analysis after the run, we offer HRM Cloud app or HRM software v3.2 (https://www.thermofisher.com/lt/en/home/technical-resources/software-downloads/high-resolution-melt-hrm-softwarev31.html) that is now also compatible with the QuantStudio 1 instrument.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Recommended maintenance schedule:
ROI/Uniformity
- Every two years (recommended)
- Always perform new Background and Dye calibrations after a ROI/Uniformity calibration.
Note: Performing a ROI/Uniformity calibration invalidates all other calibrations.
Background
- Every two years (recommended)
- Background calibration can also be performed, as needed, to check for contamination (depends on usage and laboratory conditions) and to obtain the most accurate data for the removal of background fluorescence.
Note: Performing a Background calibration does not invalidate any other calibration.
Pure dye
- Every two years (recommended)
- During a Dye calibration, only the dyes on the given spectral calibration plate are calibrated.
Note: Performing a Dye calibration for a given dye plate does not invalidate any other calibration.
RNaseP instrument verification
- After installing or moving the instrument
- After performing instrument calibrations
- As needed to confirm instrument performance
Perform an instrument self-test
- Every month
Check computer disk space. If necessary, archive or back up your experiment files and instrument settings
- Weekly
Power on/off the computer controlling the instrument
- Weekly
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
The QuantStudio 1, 3, and 5 instruments take up lesser space on the lab bench and come with one fixed block configuration. The interchangeable block systems (QuantStudio 6 and 7 Pro and the QuantStudio 6 and 7 Flex instruments) have a larger footprint but also have additional features and benefits. See more information at thermofisher.com/qpcrsystems. The QuantStudio 1 instrument is an affordable instrument that offers the most essential features for qPCR analysis: 3 color detection and the most commonly used 96-well, 0.2 mL block. It offers the capability to have Wi-Fi connection and use of the instrument with guest, user, and administrator accounts.
The main factors to consider when selecting an instrument is the multiplex reaction capacity, i.e., number of colors detected, block format needed, and if the blocks need to be interchangeable. Please see the comparative table on page 36 in this brochure (https://assets.thermofisher.com/TFS-Assets/BID/brochures/qpcr-pcr-solutions-brochure.pdf).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.