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查看更多产品信息 QuantStudio™ 3D Digital PCR Chips - FAQs (4485507)
36 个常见问题解答
If you include all of the necessary dilution factors into the AnalysisSuite software, the software will give you the copies/µL in your stock. There are 2 dilutions you need to take into account. The first is the dilution of the sample in the reaction. The second is the dilution of the stock that you make before adding it to the digital PCR reaction.
For example, say you want to add 1 µL of a sample that has been diluted 1:10 from the stock. If you add 1 µL of your sample to a 16 µL (final volume) reaction, the dilution factor of the sample is 1:16 or 1/16 = 0.0625. Since the stock has also been diluted 1:10 (0.1), you also need to factor this in. The final dilution factor to enter into the software is 0.0625 x 0.1 = 0.00625 (1:160). As you can see below, you can use either annotation to indicate the dilution factor in the AnalysisSuite software. If you enter that value into the “Dilution” column, the software will give you the copies/µL in your starting material (stock).
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
AnalysisSuite software is optimized for Google Chrome v4.0 or later. Firefox also works, but Internet Explorer is not recommended.
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Please follow the procedures starting in "Chapter A: Maintenance" of the QuantStudio 3D Digital PCR System User Guide (https://tools.thermofisher.com/content/sfs/manuals/MAN0007720.pdf) to update the instrument firmware.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
This will depend on the concentration range of the sample used. The QuantStudio 3D Digital PCR System has application- and concentration-specific requirements for technical replicates. At a low concentration, more replicates will be needed to have confidence in the data. Also, due to the nature of the application, replicates would be needed when studying rare mutations.
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The current QuantStudio 3D Digital PCR Chip allows for one sample per chip; however, duplexing allows analysis of two targets per chip.
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Please follow the procedures described starting in Appendix B of the QuantStudio 3D Digital PCR System User Guide (http://tools.thermofisher.com/content/sfs/manuals/MAN0007720.pdf) to connect the instrument to a network.
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The QuantStudio 3D Digital PCR System can read the digital chip in less than 1 minute. In fact, if you are running multiple chips, you can run the next chip in the series when the instrument touch screen displays the Analyzing Chip screen.
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After thermal cycling, the QuantStudio 3D Digital PCR 20K Chips can remain on the sample block for up to 24 hours. Read the QuantStudio 3D Digital PCR 20K Chips within 1 hour after you remove them from the GeneAmp 9700 PCR System sample block.
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Using the Chip Carrier on the GeneAmp 9700 Dual Flat Block, up to 24 QuantStudio 3D Digital Chips can be run at one time. Please refer to Chapter 3 in the QuantStudio 3D Digital PCR System User Guide (http://tools.thermofisher.com/content/sfs/manuals/MAN0007720.pdf) for more details on loading the chips for thermal cycling.
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Yes. Never handle QuantStudio 3D Digital PCR 20K Chips or Chip Case Lids without gloves. Oils from your hands can contaminate the components and interfere with thermal cycling and imaging. For more instructions, please see the guidelines in Chapter 2 in the QuantStudio 3D Digital PCR System User Guide (http://tools.thermofisher.com/content/sfs/manuals/MAN0007720.pdf) on loading the digital chips.
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Yes, The GeneAmp 9700 PCR System must be running firmware version 3.12 or later to thermal-cycle QuantStudio 3D Digital PCR 20K Chips. The firmware version of your GeneAmp 9700 PCR System is shown on the main display after you first turn on the instrument. If needed, download and upgrade the firmware.
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The system is meant to be used with the QuantStudio 3D Digital Master Mix only.
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The QuantStudio 3D instrument only performs the imaging and primary analysis of the digital chips. The chips themselves must be cycled offline on a Dual Flat Block GeneAmp 9700 PCR System.
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The QuantStudio 3D Digital PCR Chips are meant to be used on a Dual Flat Block Proflex or GeneAmp 9700 PCR System.
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The UV glue is usable for 3 months after first opening, when stored properly. Keep it stored in an amber bag while using it on the bench, and in an amber bag in a dark place between uses.
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The Tilt Base inclines the GeneAmp 9700 PCR System sample block by 11 degrees C, the angle required for optimal performance of the QuantStudio 3D Digital PCR 20K Chips.
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Use all of the immersion fluid within 60 minutes of uncapping the syringe. Once a syringe is opened, you cannot reattach the cap for later use.
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The QuantStudio 3D Digital PCR System features a filter set that is optimized for the FAM, VIC, and ROX dyes available from Life Technologies.
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The QuantStudio 3D Digital PCR 20K Chip is a 10 mm2 high-density reaction plate that consists of a single array of 20,000 reaction wells, where each well can accommodate an 865 pL reaction. Hydrophobic coatings on the chip surface enable loading and isolation of independent PCR reactions within the reaction wells. Please see an image of the chip in the QuantStudio 3D Digital PCR System User Guide, found here:
https://assets.thermofisher.com/TFS-Assets/LSG/manuals/MAN0007720.pdf
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
For instrument specifications, please see Appendix D, “Specifications and Layout”, in the QuantStudio 3D Digital PCR System User Guide for more details at http://tools.thermofisher.com/content/sfs/manuals/MAN0007720.pdf.
Find additional tips, troubleshooting help, and resources within our Real-Time PCR and Digital PCR Instruments Support Center.
Digital PCR can be used to perform rare allele detection, absolute quantitation of gene expression, viral titering, and absolute quantitation of next-generation sequencing libraries.
The QuantStudio 12K Flex system with OpenArray block and the QuantStudio 3D system are designed for digital PCR applications.
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Yes, there is an option to export the Chip data as .xml on the Export tab in AnalysisSuite software v3.0.
The Poisson Plus algorithm corrects for well-to-well load volume variation on a per Chip basis. This becomes important at higher target concentrations.
You should use the Poisson Plus algorithm for projects that contain QuantStudio 3D Chips with target quantities >2,000 copies/µL.
Yes, please use Cat. No. A27322.
At low concentrations, there is no impact on the data. At higher concentrations (>2,000 copies/µL), there is a slight impact. Using the Poisson Plus algorithm will correct for well-to-well volume variation.
No significant difference has been measured.
Yes, internal studies have shown comparable performance between v1 and v2 Chips.
Yes, Firmware v3.0 will work with all versions of Chips and Lids.
No. No matter which version of Chip Lid is being used, v2 Chips require Firmware v3.0 for proper data analysis.
We recommend that you upgrade the firmware to v3.0 in order to read the 2D barcode; otherwise the .eds files will not have a Chip ID.
No, the v2 Master Mix can only be used with the v2 Chips.
No, the current v1 Master Mix can only be used with the older Chips.
Yes, the current Chips will work with the new v2 Chip Lids.
Yes, the current Lids will work on the new v2 Chips.