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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.
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Tab 01 / DreamPrep™ NGS Solution
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DreamPrep NGS combines the Fluent® Automation Workstation, Infinite® F Nano+ plate reader and Tecan library preparation kits to provide sequencing-ready libraries with unprecedented speed and accuracy. This groundbreaking approach offers preparation, quantification, normalization and pooling of NGS libraries in one automated workflow.
Tecan’s innovative NuQuant® library quantification method uses the integrated Infinite reader to enable fully automated QC without sample loss in under six minutes, eliminating the need for slow and costly qPCR- or capillary electrophoresis-based QC.
Each DreamPrep NGS workstation is supplied with example methods* for Tecan reagent kits, including AnyDeplete™ mediated transcript depletion of unwanted reads from sequencing data.
* 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.
Rapid advances in technology have made NGS a reliable and simple option for genomic assays, but library preparation remains a challenge for high throughput applications. The speed, accuracy and precision of the Fluent platform makes it ideally suited to preparing NGS libraries, reducing process variability and ensuring greater reproducibility between operators and over time. The system’s three independent, task-specific arms operate in parallel to provide fast, efficient processing. Its built-in touchscreen interface guides users through daily tasks and can be easily customized to suit your laboratory and workflow needs for straightforward operation. Together with integrated user management and Method Approval functions, this ensures secure protocols and optimal performance.
On-deck thrmocycling for longer walkaway runs
Integrated plate reader for QC without sample loss
Magnetic separation for bead clean-ups
Prevent unauthorized changes to validated protocols with the Method Approval function.
Enhanced productivity with Tecan consumables
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Tab 02 / 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.
Show moreThe Universal Plus mRNA-Seq library preparation kit features a robust end-to-end workflow with a wide input range for analysis of coding transcripts. Universal Plus mRNA libraries can be prepared, quantified, normalized and pooled in less than nine hours on the Fluent® Automation Workstation, compared to more than 14 hours for manual or automated library preparation and qPCR quantification with other kits.
Key features of this mRNA-Seq library preparation kit include:
Trio RNA-Seq offers a complete streamlined library preparation solution that combines three core technologies to capture rare transcripts and low abundance pathogens. Trio’s proven amplification technology (single primer isothermal amplification, SPIA®) generates more molecules for library preparation compared to standard RNA-Seq workflows, preserving precious biological information from limited samples.
Key features of this RNA-Seq library preparation kit include:
The Celero EZ DNA-Seq library preparation kit is a simple, single tube, addition-only workflow with just one post-amplification bead purification step. Celero DNA libraries can be prepared, quantified, normalized and pooled in less than four hours on the Fluent® Automation Workstation.
Key highlights of this DNA-Seq library preparation kit include:
*NuQuant technology available with Plate A (92 UDI indexes) only.
The Ovation Ultralow System V2 is a DNA-Seq library preparation kit for ultra-low inputs. This kit includes our proprietary DimerFree® technology, which eliminates excess adaptor dimers to get more informative reads from your sequencing data. Ultralow V2 DNA libraries can be prepared in less than three hours on the Fluent® Automation Workstation.
Key features include:
Allegro Targeted Genotyping provides a fast, scalable, cost-effective approach for targeted sequencing of a wide variety of organisms using next generation sequencing. Allegro uses the patented Single Primer Enrichment Technology (SPET) method to specifically target regions of interest, providing information-rich sequencing data by capturing a SNP- or genotype-specific data point for every on-target sequencing read. The result is unparalleled sequencing efficiency, leading to rapid scalability and the lowest available cost per data point.
Key features
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.
Sequencing-verified, flexible protocols for Ion AmpliSeq™ library preparation, developed in collaboration with Thermo Fisher Scientific.
Show moreThe 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 03 / Qualified NGS protocols
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| Supplier | Kit | Freedom EVO® NGS workstation | DreamPrep™ NGS | |||
| Tecan | ||||||
| DNA-Seq | Celero™ DNA-Seq | |||||
| Celero™ EZ DNA-Seq | ||||||
| Ovation® Ultralow System V2 | ||||||
| RNA-Seq | Universal Plus™ mRNA-Seq | |||||
| Universal Plus Total RNA-Seq | ||||||
| Trio RNA-Seq™ | ||||||
| Ovation SoLo RNA-Seq | ||||||
| Targeted genotyping | Allegro® Targeted Genotyping V2 | |||||
| Illumina | ||||||
| DNA-Seq | Illumina DNA Prep | |||||
| Nextera XT | ||||||
| TruSeq® Nano DNA | ||||||
| TruSeq DNA PCR-Free | ||||||
| RNA-Seq | COVIDSeq Test *NEW* | Illumina qualification in progress | ||||
| TruSeq RNA Exome | ||||||
| TruSeq Stranded mRNA | ||||||
| TruSeq Stranded Total RNA | ||||||
| Forensics | ForenSeq® DNA Signature Prep | |||||
| Targeted enrichment | 16S Metagenomic Sequencing | |||||
| Nextera DNA Exome | ||||||
| Nextera Flex for Enrichment | ||||||
| Nextera Rapid Capture Enrichment | ||||||
| TruSeq Custom Amplicon Low Input | ||||||
| TruSight® Cancer | ||||||
| TruSight Cardio | ||||||
| TruSight Inherited Disease Sequencing Panel | ||||||
| TruSight One sequencing Panel Series | ||||||
| TruSight Rapid Capture | ||||||
| HLA typing | TruSight HLA v2 Sequencing Panel | |||||
| NEB | ||||||
| DNA-Seq | NEBNext® Ultra™ II DNA | |||||
| RNA-Seq | NEBNext Ultra II Directional RNA Poly(A) mRNA Isolation | |||||
| NEBNext Ultra II Directional RNA rRNA depletion | ||||||
| QIAGEN | ||||||
| DNA-Seq | QIAseq® FX DNA | |||||
| RNA-Seq | QIAseq Targeted RNA Panels | |||||
| Targeted enrichment | QIAseq Targeted DNA Panels | |||||
| Roche | ||||||
| DNA-Seq | KAPA® HyperPlus | |||||
| KAPA HyperPrep | ||||||
| KAPA HTP Library Prep | ||||||
| RNA-Seq | KAPA RNA HyperPrep with RiboErase (HMR) | |||||
| RNA HyperPrep with mRNA Capture | ||||||
| Targeted enrichment | SeqCap EZ | |||||
| Agilent | ||||||
| Targeted enrichment | Clarigo | |||||
| SureSelect® QXT Target Enrichment | ||||||
| SureSelectXT Target Enrichment | ||||||
| Haloplex® HS Target Enrichment System | ||||||
| Bioo Scientific | ||||||
| RNA-Seq | NEXTflex® Small RNA-Seq Kit v3 | |||||
| Thermo Fisher Scientific | ||||||
| Targeted enrichment | Ion AmpliSeq® Library 2.0 | |||||
| Ion Xpress® | ||||||
| Oncomine™ Comprehensive Assay v3M | ||||||
| GenDx | ||||||
| HLA typing | NGSgo® HLA typing | |||||
Method qualified/verified by vendor1
Method qualified/verified by Tecan1
Method custom developed2
1) Automated method developed according to the kit vendor’s published protocol. 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 (vendor qualified/verified) or by Tecan (Tecan qualified/verified).
2) Automated method developed according to specific customer need. Libraries tested by the customer.
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Tab 04 / 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 05 / Detection
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The Infinite® F Nano+ multimode reader offers absorbance and high sensitivity fluorescence reading modes in one compact instrument. It combines the high sensitivity of fluorescence assays and the speed provided by reading plate formats of up to 384 wells.
Read moreNGS 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.
Read moreDNA 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 06 / Literature
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In conjunction with the ever increasing throughput of NGS instruments and the option to multiplex more and more samples, sample preparation has become a bottleneck. Automation of NGS sample preparation helps to reduce the workload, process more samples in parallel and increase the reproducibility of library preparations, while minimizing sample processing errors.
Fluent’s groundbreaking design delivers more capacity and increased speed for genomic workflows. This innovative solution offers excellent precision, outstanding throughput and increased walkaway times. Get more done in less time, with greater confidence.
NGS DreamPrep is a complete, walkaway solution that combines the Fluent® Automation Workstation, the Infinite® F Nano+ plate reader and Tecan Genomics’ innovative library preparation kits to create sequencing-ready libraries with minimal manual intervention.
The automation of the Celero PCR Workflow with Enzymatic Fragmentation kit on the Freedom EVO NGS workstation provides a rapid, efficient and easy to run solution for the streamlined preparation of high quality sequencing-ready libraries.
Metagenomics is the study of genetic material extracted directly from environmental, veterinary or clinical samples, such as water, soil or stools....
Whole Genome Sequencing (WGS) offers the most comprehensive tool for the characterization of genomes. The automation of the Illumina Nextera DNA Flex library prep kit on the Freedom EVO NGS workstation provides a rapid, efficient and easy solution to prepare up to 96 high quality sequencing-ready libraries in a day.
Celero DNA-Seq kits with NuQuant® provide an innovative and streamlined DNA library preparation workflow for generating quantified libraries that are ready for sequencing on Illumina instruments.
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.
Targeted enrichment reduces costs and enhances the efficiency of next generation sequencing (NGS) for the identification and analysis of variations in specific regions of the genome, providing a useful tool for applications such as clinical genomics research.
Sequencing of PCR products has become a routine technique as it 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.
High quality Ion AmpliSeq libraries for inherited disease and cancer research using custom panels of up to 96 samples in less than six hours.
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. It can accommodate 500 pg to 1 μg of input DNA, sheared by either mechanical or enzyme-based methodsand prepare up to 96 libraries per run.
The automated method for TruSeq Nano DNA generates excellent quality data from as little as 200 ng – and up to 1 μg – of DNA. This allows samples with restricted DNA vailability, for example tumor biopsies, to be studied, helping to preserve precious sample material for future use. In comparison, the TruSeq DNA PCR-Free Library Preparation Kit, offers a time-saving alternative for more concentrated samples with initial quantities of between 1 and 2 μg of DNA, generating libraries with fewer coverage gaps by the elimination of PCR-induced bias, providing greater access to the genome.
Illumina’s 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.
HLA gene sequencing is a technique of growing interest, as it allows improved determination of the HLA profile of tissue donors and acceptors compared to classic techniques, such as serotypical analysis.
Illumina’s TruSeq technology delivers stranded information, identifying which DNA strands a given RNA transcript has derived from. This information ensures increased confidence in transcript annotation and raises the percentage of alignable reads, ultimately reducing the sequencing cost per sample. This application note describes automation of the TruSeq RNA library preparation kits.
The Illumina Nextera Rapid Capture Exome and Expanded Exome kits provide a fast and simple, all-in-one sample preparation and hybridization-based enrichment process for identifying coding variants in targeted resequencing studies.
Agilent's SureSelectXT 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.
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.
The Celero PCR Workflow with Enzymatic Fragmentation DNA-Seq Kit offers a simple, addition-only workflow for generating quantified libraries ready for sequencing.
The Universal Plus mRNA-Seq library preparation kit with NuQuant offers a streamlined workflow for the generation of sequencingready quantified libraries
NuQuant® is a novel library quantification method, which accurately measures molar library concentration without the need for separate fragment size analysis.
User guide for NEBNext® UltraTM II Directional RNA Library Prep Kit for Illumina® on the Freedom EVO® NGS workstation
User guide for TruSeq Custom Amplicon Low Input Kit
Prepare Ion AmpliSeq Libraries using the Tecan Freedom EVO® NGS Workstation
User guide for NEBNext Ultra II DNA Library Prep Kit
User guide for Illumina TruSeq Nano DNA and TruSeq DNA PCR-Free library preparation
User guide for Nextera XT DNA library
User guide for TruSight HLA v2 Sequencing Panel library preparation
User guide for Illumina TruSeq Stranded mRNA library preparation
User guide for Illumina TruSeq Stranded Total RNA library preparation
User guide for Illumina TruSeq RNA Access library preparation
User guide for the Illumina Nextera Rapid Capture library preparation
User Guide for Agilent SureSelect XT Target Enrichment System
User guide for NimbleGen SeqCap EZ Library SR with KAPA™ HTP Library Preparation
Research use only – not for use in diagnostic procedures.