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QUICK REFERENCE
QuantStudio
™
Catalog Number 4482593 and 4481097
Pub. No. MAN0008159 Rev. B.0
Note: For safety and biohazard guidelines, refer to the "Safety"
appendix in the QuantStudio
(Pub. no. MAN0007720). Read the Safety Data Sheets (SDSs) and
follow the handling instructions. Wear appropriate protective
eyewear, clothing, and gloves.
Operational workflow
The following figure shows the workflow for performing a single
experiment on the QuantStudio
procedures for sample and/or consumable preparation and result
analysis can vary depending on the specific experiment that you
are performing.
Set up the PCR reaction by mixing sample, master
mix, and assay(s).
Load the PCR reaction onto a QuantStudio
Digital PCR 20K Chip, apply a lid, load the assembly
with immersion fluid, then seal the loading port.
Perform the PCR using the ProFlex
System (or Dual Flat Block GeneAmp
Read the Digital PCR 20K Chip using the
QuantStudio
3D Digital PCR Instrument.
™
Review the results on the QuantStudio
Instrument touchscreen.
Store or discard the Digital PCR 20K Chip.
Analyze the data using the QuantStudio
AnalysisSuite
For Research Use Only. Not for use in diagnostic procedures.
3D Digital PCR System
3D Digital PCR System User Guide
™
3D Digital PCR System. The
™
Start
q
q
3D
™
q
2x Flat PCR
™
PCR System
®
9700).
q
q
3D
™
q
q
3D
™
Software.
™
q
Finish
Prepare the digital PCR reactions
Required materials
• Genomic or complementary DNA sample(s)
• TaqMan
Assay(s) for your experiment
®
• QuantStudio
3D Digital PCR Master Mix
™
• Gloves, powder-free, nitrile
• Laboratory-grade, lint-free wipes
• Microcentrifuge
• Permanent marker or pen
• Pipettes and tips, P10 to P1000
• Reaction tubes, DNase/RNase-free, non-stick or low-binding,
0.5-mL or 1.5-mL
• TE Buffer, 1X Molecular Biology Grade
• Vortexer
• Water, DNase-free, sterile-filtered
Prepare the reaction mix and samples
Note: The volumes of the following example protocol have been
adjusted so that two chips are run for each sample.
Table 1 PCR Reaction Mix
Material
Digital PCR Master Mix, 2X
TaqMan
Assay, 20X
®
Diluted DNA
Water
Total volume (for 1 sample/2 chips)
[1]
Volumes include 20% excess to compensate for volume loss from pipetting.
[2]
The recommended DNA volume is based upon a human gDNA sample at
10 ng/µL concentration with the target sequence present at two copies per
diploid genome. This recommended volume will vary depending upon species,
sample type, and sample concentration. For best results, we recommend
adding sufficient DNA such that the concentration of target sequence is
between 200 and 2,000 copies/µL.
1. Remove from storage and allow the QuantStudio
PCR Master Mix and TaqMan
temperature.
Volume
Stock
Final
[1]
(μL)
17.4
2X
1X
1.7
20X
1X
[2]
3.5
10 ng/μL
1 ng/μL
12.2
—
—
34.8
—
—
3D Digital
™
Assay(s) to warm at room
®
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Related Manuals for Applied Biosystems QuantStudio 3D
Summary of Contents for Applied Biosystems QuantStudio 3D
- Page 1 QUICK REFERENCE QuantStudio 3D Digital PCR System ™ Catalog Number 4482593 and 4481097 Pub. No. MAN0008159 Rev. B.0 Prepare the digital PCR reactions Note: For safety and biohazard guidelines, refer to the “Safety” appendix in the QuantStudio 3D Digital PCR System User Guide ™...
- Page 2 2. Review the concentration of your DNA samples, and prepare Load and seal the chips using a Chip Loader dilutions if necessary. WARNING! ULTRAVIOLET LIGHT HAZARD. Looking 3. Gently invert the tube of Digital PCR Master Mix 10 times (or gently vortex on low-medium speed).
- Page 3 3. Prepare a syringe containing QuantStudio 12K Flex c. Press the sample loading blade lever, then install the ™ OpenArray Immersion Fluid: QuantStudio 3D Digital PCR Sample Loading Blade to ® ™ the Chip Loader as shown. a. Remove the syringe, plunger, and tip from the packaging.
- Page 4 5. Rotate the Chip Loader arm so that the Chip Case Lid solidly b. Insert the Digital PCR 20K Chip assembly into the UV- contacts the Digital PCR 20K Chip, firmly press down for Curing Station on the Chip Loader. Push the chip into 15 seconds to ensure a tight seal, then press the Lid Nest the station until the ultraviolet light illuminates.
- Page 5 5. Open the QuantStudio 3D Digital PCR 20K Chip package, Load and seal the Digital PCR 20K Chips ™ gently grasp the chip by its sides, then place it face-up on the heated block, preheated to 40°C. WARNING! ULTRAVIOLET LIGHT HAZARD. Looking directly at a UV light source can cause serious eye damage.
- Page 6 e. Allow the Digital PCR 20K Chip to sit on the heated b. Power on the UV Stylus in the UV Stylus Stand by block for 20 seconds. twisting the cap near the power button. f. Add 10-15 drops of Immersion Fluid directly to the chip c.
- Page 7 3. Load the QuantStudio 3D Digital PCR 20K Chips into the Table 3 GeneAmp PCR System 9700 PCR Method ® ™ thermal cycler: a. Place the sealed Digital PCR 20K Chips onto the sample PCR Protocol Rxn. block so that the fill ports on the chips are positioned Speed Vol.
- Page 8 You can read another Digital PCR 20K Chip without waiting Image the Digital PCR 20K Chips for the analysis to complete. To begin the next run, load the chip and touch Start Run as described in step 3. 1. Set the destination for the imaging data: a.
-
Page 9: Analyzing The Data
Analyzing the data About data quality flags Data quality flags generated and displayed by the software are a About the results screen measure of the overall chip quality, based on well quality The QuantStudio 3D Digital PCR Instrument displays a summary ™... - Page 10 Kits, consumables, and accessories Cat. no. Description 4485507 QuantStudio 3D Digital PCR 20K Chip Kit, 12-pack ™ • QuantStudio 3D Digital PCR 20K Chip (12) ™ • QuantStudio 3D Digital PCR Chip Case Lid (12) ™ • QuantStudio 3D Digital PCR Sample Loading Blade (12) ™...
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