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From just a few samples a week to hundreds a day, automation of NGS sample preparation allows you to reproducibly create high quality libraries for reliable next-generation sequencing (NGS).
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Tab 01 / Overview
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Validation and verification-ready Tecan’s reagents and 3rd party’s kits to develop automated NGS library prep protocols to ensure excellent performance in any lab.
From plug-and-play to high to highly flexible automated solutions. Now you can walk-away with confidence.
From reagents, automation, and consumables to application expertise – we support you every step of the way.
From low to high throughput, turnkey solutions to OEM development, our comprehensive NGS library prep solutions allow scientists to save time on library prep and focus on what matters most.
We continually strive to offer innovative, accessible and always reliable solutions.
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Tab 02 / DreamPrep® NGS Solutions
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Output
Software packages included
Three configurations are available and upgradable at any time
Basic: Increase your productivity as all liquid transfers and incubations are covered by the platform
Full: Integrate Quantification and Normalizations steps in your NGS workflows by including the Infinite® 200 Pro Reader F Nano+
Software packages included
All components2 provide high performance across different library preparation kits
Enabled by the integrated plate reader and NuQuant®3 technology
For faster and more efficient workflows
Parallel movement of robotic arms increases productivity
User-friendly interface empowers your daily operations
On-deck thermal cycler enables longer walk-away times
These platforms combine the advanced technology of the Fluent® automation workstation, Infinite® 200 Pro Reader F Nano+ and Tecan NGS library preparation kits to generate sequencing-ready NGS libraries in one simple automated workflow.
Tecan’s innovative reagent core technology NuQuant3 and the integrated Infinite reader enable you to simultaneously measure your final libraries on a plate without unnecessary sample loss.
1) Number of samples processed in walk-away mode is determined by the NGS protocol
2) Ring magnet plates and POGO® adapter with Spring Cushion Technology manufactured by ALPAQUA® Engineering, LLC. Beverly, USA.
2) Thermal devices (Cold Plate Liquid-Cooled, CPLC) and ODTC® manufactured by Inheco© Industrial Heating and Cooling GmbH. Martinsried, Germany.
2) BioShake®3000-T elm manufactured by QInstruments® GmbH. Jena, Germany.
2) Infinite® 200 PRO F Nano+ Reader is a product of Tecan Trading AG, Switzerland.
3) NuQuant® is a registered trademark of Tecan Group Ltd., Männedorf, Switzerland or of Tecan Genomics, Inc., Redwood City, USA.
Example methods have not been validated. Please make a copy of these methods to customize them to accommodate and validate your workflow according to your desired intended use and laboratory protocols. Tecan makes no claims regarding the performance of the example methods.
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Tab 03 / Reagents
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Tecan has developed and optimized its own NGS library preparation reagents for a broad range of sample types. These reagents have been verified on Tecan automation platforms to give you a complete solution, from samples to readily normalized libraries.
The TruSeq Nano DNA Sample Preparation Kit enables efficient investigation of samples where the amount of DNA available is limited. The automated workflow for this kit on the Freedom EVO® NGS workstation includes simple bead-based size selection, and can be completed in less than a day with minimal hands-on time for up to 48 samples.
The TruSeq DNA PCR-Free Sample Preparation Kit enables time-saving investigation of DNA samples. The user-friendly TouchTools™ interface guides the operator through automation set-up to deliver highly reproducible, sequencing-ready DNA libraries for single read, paired-end and indexed sequencing with minimal user intervention.
The Nextera XT DNA Library Preparation Kit enables the effective construction of high quality libraries from PCR amplicons, plasmids and small genomes, using a protocol optimized for DNA inputs from as little as 1 ng.
RNA sequencing (RNA-Seq) is a powerful method for discovering, profiling and quantifying RNA transcripts. Using Illumina next generation sequencing technology, stranded information identifies the DNA strand from which a specific RNA transcript was derived, providing increased confidence in transcript annotation and visibility into antisense regulation.
Automation of the Illumina TruSeq Stranded Total RNA protocol on the Freedom EVO NGS workstation enables the conversion of total RNA into libraries of template molecules of known strand origin with minimal user intervention. The Freedom EVO NGS workstation delivers high quality libraries, with minimal risk of cross-contamination.
The TruSeq RNA Access library preparation protocol is the ideal solution for cost-effective sequencing of formalin-fixed paraffin-embedded (FFPE) archival tissue samples. The integrated enrichment steps can accommodate low starting concentrations of DNA, and the automated workflow allows the construction of up to 48 libraries in parallel, generating high quality libraries with limited user intervention and minimal risk of cross-contamination.
The automation-friendly workflow of the Nextera Rapid Capture protocol combined with the Freedom EVO NGS workstation provides a faster, more efficient solution for library preparation and exome enrichment. Using the automated Nextera Rapid Capture Exome protocol, sample preparation can be completed with minimal hands-on time in just a day and a half, generating sequencing data results the following day.
Interest in HLA gene sequencing is growing, as it offers better determination of the HLA profile of tissue donors and recipients compared to classic techniques, such as serotypical analysis.
The Illumina TruSight HLA v2.0 Sequencing Panel is a reliable and simple assay designed to generate NGS libraries for HLA profiling. Automation of the workflow on the Freedom EVO NGS workstation allows sequencing of 11 HLA loci for up to 24 samples, minimizing the risk of cross-contamination and human errors during library preparation.
Amplicon sequencing is a targeted method for identification and analysis of variations in specific regions of the genome. Sequencing of PCR products offers cost-effective variant identification and characterization. Automation of the TruSeq Custom Amplicon Low Input library preparation workflow on the Freedom EVO NGS workstation provides a fast and efficient solution. This setup enables generation of high quality libraries from low input samples – starting with as little as 10 ng of genomic DNA.
The NEBNext Ultra II Directional RNA Library Prep Kit for Illumina uses the ‘dUTP` method to produce excellent yields and high quality directional libraries even from low quality RNA. Up to 96 libraries can be prepared at a time from a broad range of input material, 10 ng - 1 µg total RNA for poly(A) mRNA enrichment, 5 ng - 1 µg for rRNA depletion.
The NEBNext Ultra II DNA library prep workflow combines the end repair and dA-tailing steps with minimal clean-¬up to reduce sample preparation time. Up to 96 libraries can be prepared per run, and multiplexed using, for example, the NEBNext Multiplex Oligos for Illumina.
This protocol allows highly reproducible library preparation from a wide range of input DNA concentrations from 500 pg to 1 μg, as well as FFPE samples, offering flexible processing of 1 to 96 samples with minimal user intervention.
Agilent’s SureSelect XT Target Enrichment System provides a complete portfolio of exome kits for humans and other species, as well as custom kits for both DNA and RNA capture.
Click here for an application note describing an automated protocol for parallel processing of up to 48 samples using the SureSelect XT Target Enrichment System on the Freedom EVO NGS workstation.
Roche NimbleGen’s SeqCap EZ Library is a solution-based capture system enabling enrichment of the whole exome or custom regions of interest from DNA sequencing libraries. The SeqCap EZ protocol covers exome designs, gene panels and custom content designs, according to Roche NimbleGen specifications.
Click here for an application note describing an automated protocol for NimbleGen SeqCap EZ with KAPA™ HTP Library Preparation in combination with NimbleGen SeqCap Adapter Kits A and/or B. The protocol allows preparation of up to 96 libraries, and capture of up to 24 library pools.
The Ion AmpliSeq Library Kit 2.0 enables rapid NGS library preparation for 12-25,000-plex PCR in a single well.
Automated library preparation of up to 96 samples can be performed on the Freedom EVO NGS workstation in less than six hours using 10 ng of DNA or RNA. The TouchTools operator interface minimizes hands-on time and training requirements, guiding users through every stage of protocol selection and worktable set-up.
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Tab 04 / Qualified NGS protocols
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Method available: |
Automated method developed according to the kit vendor’s published protocol. Protocol was tested using water and/or reagents with Tecan instruments. |
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Method qualification in progress: |
Automated method developed according to the kit vendor’s published protocol. It is currently being tested with reagents and commercial samples by the vendor. |
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Method qualified: V T C |
Automated method developed according to the kit vendor’s published protocol. Method was qualified using kit vendor’s guidelines, reagents and commercial samples. Libraries prepared with these methods meet the kit vendor’s specifications and/or perform comparably to those prepared manually. Sequenced libraries were assessed either by the vendor (V vendor qualified), by Tecan (T Tecan qualified) or by the customer (C customer qualified). |
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Method custom developed: |
Automated method developed according to specific customer need. Libraries tested by the customer. |
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Tab 05 / Freedom® Evo NGS workstation
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There are so many things that you have to get right to create high quality libraries for reliable next generation sequencing, so why have unnecessary complexity in your liquid handling?
Automation of NGS sample preparation on the Freedom EVO NGS workstation increases throughput and reproducibility, guiding the user through each step of protocol selection and worktable set-up with the TouchTools™ operator interface.
The Freedom EVO NGS workstation is equipped with innovative air displacement pipetting technology, using eight pipetting channels (1) to allow precise pipetting from 1,000 µl down to 0.5 µl. The workstation includes three Inheco CPAC devices (2) to keep reagents cool and provide optimal conditions for the enzymatic steps, an Inheco Thermoshake heated shaker (3), a 96-position Alpaqua 96S Super Magnet Plate magnetic plate separator (4) and a RoMa Arm (5) for efficient bead clean-up. In addition, the compact worktable provides storage space for up to 12 tip boxes (6), allowing longer unattended runs.
Precision: The Freedom EVO’s precision automation ensures the reproducibility necessary for high quality results, with a range of qualified sequencing-verified protocols available.
Flexibility: Don’t be constrained by batching requirements. Whether you need to process 6, 13, 19, 40 or even 96 samples, only run what you need, saving on reagents.
Simplicity: You don’t need to be an automation expert. With the simple touchscreen interface, you can get started right away, set up your runs quickly and be confident in your workflows.
The TouchTools application starter screen lets the operator select the desired protocol with the touch of a button. Step-by-step user instructions help to avoid potential errors during worktable set-up to ensure the correct initial conditions. A selection confirmation screen then provides a summary as a final check before starting the run, making it easy for anyone in the lab to use the instrument.
The Leiden University Medical Center has integrated the Freedom EVO® NGS workstation into its molecular diagnostic testing and clinical research workflows. Taking advantage of this preconfigured solution, the center was quickly able to commission the platform for routine testing, while still having the flexibility to develop new protocols and conduct research studies.
The University of North Carolina (UNC) has automated its major sample preparation protocols for next generation sequencing on a Freedom EVO® workstation, enabling more efficient processing.
The GeT genomics facility near Toulouse has been developing a series of automated NGS protocols in collaboration with Tecan – including Illumina’s TruSeq® DNA and RNA kits, as well as Bioo Scientific’s NEXTflex™ PCR-Free Modules – on two Freedom EVO® workstations, helping to improve throughput and reliability for a wide range of projects.
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Tab 06 / Detection
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NGS library quantification is a crucial step for a successful sequencing run. Accurately determining the number of sequenceable molecules is highly significant for new high capacity sequencers, such as the NovaSeq system.
Tecan NuQuant® is a novel library quantification method that accurately measures molar library concentration without the need for separate fragment size analysis. This proprietary method uses a fluorescent label that is incorporated into the library molecules during library preparation. NuQuant is independent of library size, allowing library molarity to be directly measured using fluorometers, such as the Infinite F Nano+ multimode reader.
NuQuant library quantification on the Infinite F Nano+ plate reader eliminates the need for additional manual qPCR or capillary electrophoresis steps, saving significant time, resources and valuable library material.
After final purification of the DNA libraries in a 96-well plate on the Fluent ® Automation Workstation, the fluorescence of the libraries and standards is measured on the Infinite plate reader. The molarity determined for each library can then be used for balanced pooling and subsequent sequencing on Illumina instruments.
DNA quantification can be achieved by measuring an aliquot with a fluorescent dye, such as PicoGreen® or QuantiFluor™, or by measuring the absorbance of the entire DNA sample. DNA quantification and normalization workflows typically include the following steps
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Tab 07 / Literature
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Research use only – not for use in diagnostic procedures.