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NGS sample preparation

Automated library preparation, quality control and pooling

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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System highlights

  • DNA-Seq libraries, normalized and ready to sequence, in less than four hours
  • mRNA-Seq libraries, quantified and ready to sequence in nine hours instead of 14
  • Integrated, full library QC in less than six minutes without sample loss
  • Addition-only workflows for DNA-Seq, plus minimal intervention for RNA-Seq
  • Single point of contact for automation, reagents, consumables, application support, etc.
  • Intuitive, touchscreen-based user interface that can be customized to your needs

 

* 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

Intuition built right in

Simply faster and more precise

Save reagents and samples

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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Tab 03 / Qualified NGS protocols
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    NGS_Dreamprep-FreedomEvo_300x300

 

NGS_Dreamprep-Fluent_300x300

 

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.

TouchTools operator interface

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.

Tecan Journal Articles

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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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.




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Tab 06 / Literature
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Brochures

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.

Application Notes

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 Guides

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