The QuantStudio™ 12K Flex Real-Time PCR System helps streamline targeted discovery and validation, pathogen detection, and biomarker identification and screening, at a scale previously unattainable. Achieve the highest sample throughput for mid-density gene expression analysis - in a single day, one user can generate >42,000 gene expression data points without the use of robotics.

  • More replicates—each plate generates 2,600 data points, enabling you to include the technical replicates needed for better statistical power
  • Simple workflows—each QuantStudio™ 12K Flex OpenArray® Plate, encased in an alloy bottom for easy handling, comes preloaded with the assays of your choice- just add sample and master mix, cycle, and image
  • Experimental flexibility—specify the inventoried TaqMan® Gene Expression Assays you need, and select the OpenArray® Plate format that best meets your project needs
  • Reliable and economical—TaqMan® MGB probe chemistry in 33 nL reactions reduces reagent usage

TaqMan® OpenArray® gene expression workflow on QuantStudio™ 12K Flex System

Isolate and purify total RNA.

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RNA extraction for Real-Time PCR
One key variable to the success of any real-time PCR experiment is optimal RNA extraction. The one issue that all RNA isolation methods have, however, is the inability to completely remove genomic DNA contamination from the RNA sample. DNA removal is especially critical for RT-PCR applications because DNA can serve as a template during the PCR portion of the process, resulting in false positives, background, etc. Ambion® offers products to simplify your RNA real-time PCR experiments and minimize the risk of genomic DNA contamination to help make your data more quantitative.

Select inventoried TaqMan® Assays and configure the OpenArray® plate layout using the downloadable workbook tool. QuantStudio™ TaqMan® OpenArray® Real-Time PCR Plates are delivered with assays dried down in the through-holes you specify. Mix DNA samples with master mix in OpenArray® 384-well sample plate.

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TaqMan® OpenArray® Real-Time PCR Plates
TaqMan® OpenArray® Real-Time PCR Plates deliver world-class TaqMan® Assays in a more flexible, high-throughput format. Choose from five customizable plate formats and over 61,000 inventoried TaqMan® Gene Expression Assays. Plates arrive with your assays  pre-loaded in the 3,072 through-holes on the OpenArray® Plates.

Configure a Gene Expression OpenArray® Plate using pre-designed TaqMan® Gene Expression Assays.
Configure a Gene Expression OpenArray® Plate using inventoried, pre-designed, and custom TaqMan® Gene Expression Assays.

OpenArray® Master Mix, Loader Tips & other plastics:
QuantStudio™ 12K Flex OpenArray® Gene Expression Starter Kit

Reverse transcription: Choosing the correct reverse transcriptase (RT) for cDNA synthesis is critical to obtaining high yields of cDNA to improve the range of your real-time RT-PCR assays. The SuperScript® VILO™ cDNA Synthesis Kit is designed to increase cDNA yields and improve the range of your qRT-PCR assays. You obtain the same relative representation in your cDNA, irrespective regardless of the abundance of your gene of interest.

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SuperScript® VILO™ cDNA Synthesis Kit – 50 reactions
SuperScript® VILO™ cDNA Synthesis Kit – 250 reactions

Pre-amplification: Pre-amplification is an optional and intermediate amplification step that makes it possible to perform gene expression analysis starting with a limited quantity of RNA. Pre-amplification enables you to stretch your precious sample into many more real-time PCR reactions.

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TaqMan® PreAmp Master Mix

Prepare your sample mix: Mix cDNA or the product of your pre-amplification step with TaqMan® OpenArray® Genotyping Master Mix in a TaqMan® OpenArray® 384-well sample plate.

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TaqMan® OpenArray® Real-Time Master Mix (1.5 mL, 2X)
TaqMan® OpenArray® Real-Time Master Mix (5 mL, 2X)
OpenArray® 384-Well Sample Plates
OpenArray® 384-Well Barcoded Sample Plates

Load your sample mixes onto the TaqMan® OpenArray® Real-Time PCR Plate using the OpenArray® AccuFill™ System. Next, insert the TaqMan® OpenArray® Real-Time PCR Plate into the OpenArray® case filled with immersion fluid, and seal with adhesive. Run the reactions and collect your data using the OpenArray® Real-Time PCR Instrument.

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TaqMan® OpenArray® Real-Time PCR Master Mix
QuantStudio™ 12K Flex OpenArray® AccuFill™ System
The OpenArray® AccuFill™ System loads the sample mix from a 384‐well plate onto a TaqMan® OpenArray® Real-Time PCR plate with minimal human intervention.
OpenArray® Sample Tracker Software
OpenArray® Sample Tracker Software integrates the sample i nformation from a 384‐well plate file with the assay information contained in the plate setup file.
OpenArray® AccuFill System Tips
OpenArray® AccuFill™ System Tips, 10 pack

Load sample mixes onto TaqMan® OpenArray® Real-Time PCR Plate using the QuantStudio 12K Flex OpenArray® AccuFill™ System. The plate is encased with an alloy bottom for easy handling. Simply apply the adhesive lid and fill with immersion fluid. Perform a real-time plate read using the QuantStudio™ 12K Flex system.

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QuantStudio™ 12K Flex Real-Time PCR System with OpenArray® Block
The QuantStudio™ 12K Flex Real-Time PCR Instrument and QuantStudio™ 12K Flex OpenArray® AccuFill™ System combination can process hundreds to thousands of samples per day, enabling mid-density, high-throughput results with ease of use and without robotics.

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Acquire, analyze, and manage data quickly and easily. Amplification curves are presented for review, and data is easily manipulated to calculate Ct and Tm, as well as ΔCt and ΔΔCt measurements.

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ExpressionSuite Software
Applied Biosystems® ExpressionSuite Software is a free, easy to use data analysis tool that utilizes the Comparative Cτ (ddCτ) method to rapidly and accurately quantify relative gene expression across a large number of genes and samples.

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Literature

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