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NGS Technology powered innovation

A range of proprietary technologies, all innovatively designed to simplify and accelerate NGS workflows, increase detection sensitivity and maximize informative sequencing and gene expression data from any sample.
Overview




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Tab 01 / Overview
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Tecan’s NGS products incorporate a range of proprietary innovations, all designed to simplify and accelerate workflows, increase detection sensitivity, address common limitations and maximize sequencing of informative reads and gene expression data from any sample.

It is these technologies that set Tecan's kits apart from the norm: able to continue working at lower input concentrations or retrieve data from poor quality and degraded samples, incorporating time and cost-saving features that streamline workflows, eliminate steps and ensure only the desired reads end up on the sequencer so that data is cleaner, and analysis is quicker.

 

 

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Find out more about our core enabling technologies below:

  • Single Primer Isothermal Amplification (SPIA) – robust, high sensitivity cDNA amplification. Incorporated in RNA library preparation kits for sensitive detection of rare transcripts from challenging samples.
  • NuQuant®– Accurate quantification of NGS libraries determine the molar concentration of your library in minutes with a simple fluorescent measurement. 
  • AnyDeplete® – remove any unwanted, high abundance transcripts from your library prior to sequencing.
  • Single Primer Enrichment Technology (SPET) - Enrich your final library with only targeted regions of DNA. Focus on only the sequence you are interested in for targeted re-sequencing, Genotyping, SNP discovery and any other target specific application.
  • Unique Dual Index (UDI) adaptors - Allow you to take full advantage of the increased sequencing capacity of Illumina sequencing instruments and avoid the problems of index hopping with up to 384 pre-plated UDI adaptors.
  • DimerFree® – eliminate primer dimers in the final library.




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Tab 02 / SPIA
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SPIA – High sensitivity detection and amplification of RNA from any sample type

Single Primer Isothermal Amplification (SPIA) is an unbiased amplification technique which preserves biological information from your samples. A rapid, user friendly protocol that is compatible with challenging sample sources, low or degraded target RNA, and tolerant of inhibitors.

SPIA amplification technology is a well-established, highly published, and peer-reviewed method for generating cDNA libraries from RNA samples with a level of sensitivity far below the detection threshold of other techniques.

Benefits

SPIA is a highly reproducible method of isothermal amplification compatible with a broad range of samples and workflows.

  • Works with RNA from all sample types–Liquid biopsies, FFPE, LCM, cell lines, tissues.
  • Increased sensitivity - able to detect extreme low copy transcripts at levels below the detection threshold of other methods.
  • Yields over a microgram of high-quality sequence ready cDNA from even the lowest quality, degraded samples.
  • Unbiased amplification - preserving the biological information contained in the samples.
  • Automated workflows available on a variety of liquid handling platforms.

SPIA Workflow

In Step 1, amplified cDNA is generated with priming at the 3’ end as well as randomly throughout the transcriptome.

In Step 2, SPIA primers are added and amplification runs to completion at an isothermal temperature and without requiring optimization.

In Step 3, the finished, amplified cDNA library is ready to be used in any normal application such as sequencing, qPCR, microarray or stable archiving of the sample.

Sensitive, robust RNA amplification

The figure demonstrates the performance of SPIA with a mix of low quality, degraded, and low concentration RNA.

As can be seen from the RNA traces, the quality of the RNA is very low (RIN<3).

Yet in each case the SPIA technology was used to amplify the sample creating a viable cDNA library.

Nasal swab SPIA RIN

Find this technology in:

RNA-SeqRevelo™ RNA-Seq library preparation kit

Revelo RNA-seq library preparation solutions combine proprietary technologies to provide a streamlined solution for whole transcriptome RNA-seq from low-input or poor quality samples for rare transcript deletion and unbiased pathogen discovery.

DNA-SeqCrescendo cDNA™ Synthesis for qPCR

Robust solution for enhanced qPCR detection from low input and degraded samples Quantitative PCR (qPCR) is a sensitive method that allows researchers to detect and quantify nucleic acids in a starting material.

RNA-Seq Trio RNA-Seq Library Preparation Kit

Unique RNA-Seq library prep combines three proprietary technologies to provide a streamlined solution for whole transcriptome RNA-Seq library preparation from low input and poor quality samples for rare transcript detection and unbiased pathogen discovery.

RNA-Seq Ovation® RNA-Seq System V2

Single Primer Isothermal Amplification (SPIA) technology provides industry leading whole transcriptome reverse transcription and cDNA amplification for low input and degraded RNA samples. The amplified cDNA can be used in a range of applications from NGS library prep and qPCR to sample archiving.




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Tab 03 / NuQuant
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NuQuant®: 1-step library quantification. 6 min. qPCR accuracy.

Integrated NGS library prep and quantification in a single workflow. Accurate library quantification in just minutes.

Benefits

  • Scale your assay throughput – compatible with plate readers, you can automate your workflow from sample preparation all the way to library quantification.
  • Save time and cost – No need to measure the amount (qPCR) and size (Fragment Analyzer) as NuQuant directly measures molarity in one quick assay.
  • Never throw away your precious samples – Libraries labelled with NuQuant are compatible with Illumina® platforms and do not interfere with cluster formation and sequence detection. This allows for sample to sequencer with no sample throwaway.

Optimized cluster generation is critical to the success of sequencing runs on any next-generation platform, and cluster density is directly driven by the amount of library DNA loaded onto the flow cell.

  • Too much DNA results in over-clustering, lower Q30 scores, poor sequencing resolution, less readable data, and costly wasted reads.
  • Too little DNA leads to under-clustering, with wasted space on the flow cell and less readable data.

Both situations have the same outcome: costly and time-consuming re-sequencing of failed runs.

The key to optimized cluster generation is accurate library quantification.

For years, the gold standard for library quantification has been qPCR – the only method with sufficient accuracy to ensure optimal flow cell loading. However, qPCR is a complex procedure that takes upwards of 90 minutes to get results. By contrast, NuQuant provides results in just 6 minutes, with qPCR levels of accuracy, and no dilutions, requiring only that the plate be read on any standard lab fluorimeter.

Comparison of library quantification methods

Comparison of library quantification methods

NuQuant is the only technology that can match the accuracy of qPCR, measuring only the sequenceable fragments and ignoring any other nucleic acids.

NuQuant Workflow

NuQuant is a proprietary method that does not require additional reagents or secondary library analysis methods, making it fast and simple.

Library Quantification Workflow Comparison

Library Quantification Workflow Comparison

With NuQuant library quantification, a specific number of fluorescent labels are incorporated into the library molecules during library preparation.

As each library molecule has an equivalent number of labels incorporated, regardless of the size of the library fragment, the library molar concentration of 96 samples can be directly measured in less than 10 minutes using Infinite® or any standard plate reader. Alternatively, samples can be individually measured using Qubit® in around 6 minutes per sample.

NuQuant results are equivalent and correlated to gold standards

Comparison of NuQuant vs. qPCR vs. Qubit

NuQuant and qPCR results are comparable and functionally equivalent.

Dilution and pooling calculations using either of these two data sets would yield equivalent results, and both are clearly very different from the results obtained from Qubit.

Correlation between NuQuant and iSeq100

The correlation between NuQuant and iSeq100 (the gold standard for library quantification) over nine library pool samples clearly demonstrates the accuracy of NuQuant.

Find this technology in:

DNA-SeqCelero™ DNA-Seq Library Preparation Kit

Simple DNA-Seq library preparation kit that enables library construction in three steps. NuQuant library quantification technology included providing accurate library molarity in seconds.

DNA-SeqCelero™ EZ DNA-Seq Library Preparation Kit

Are you looking for a robust, time-saving DNA-Seq workflow? Celero EZ DNA-Seq Library preparation kit is a simple 3-step workflow integrated with NuQuant library quantification that provides a flexible solution for high-throughput DNA-Seq.

RNA-SeqRevelo™ RNA-Seq library preparation kit

Revelo RNA-seq library preparation solutions combine proprietary technologies to provide a streamlined solution for whole transcriptome RNA-seq from low-input or poor quality samples for rare transcript deletion and unbiased pathogen discovery.

RNA-Seq Universal Plus mRNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for mRNA-Seq library preparation for sequencing on Illumina sequencers. This kit features a broad input range, UDI, and simple library quantification with NuQuant.

RNA-Seq Universal Plus Total RNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for total RNA-Seq library preparation for sequencing on Illumina sequencers. This kit features broad input range, UDI adaptors, rRNA depletion with AnyDeplete and simple library quantification with NuQuant.




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Tab 04 / Anydeplete
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AnyDeplete® – Targeted depletion of unwanted transcripts

AnyDeplete maximizes the informative sequence reads in whole transcriptome data by eliminating the abundant unwanted transcripts. Uncover more of the hidden gems hiding in data and reduce sequencing costs at the same time with Anydeplete.

AnyDeplete designs are customizable for any sample type and any species, with over 15 standard designs covering the most common model species plus humans, and hundreds of custom designs.

Benefits

AnyDeplete effectively eliminates unwanted rRNA transcripts from the library, leaving behind only the desired reads; yielding more informative data from every experiment.

  • Efficient rRNA depletion regardless of input amount, from as little as a single cell to a microgram of total RNA.
  • Completely customizable to remove any unwanted sequence in your library. High quality cost-effective data from any sample source.
  • Unlike other methods, transcript depletion occurs after library construction. No reduction of input RNA concentration and no bias in the final library.
  • AnyDeplete effectively eliminates rRNA.

AnyDeplete Workflow

AnyDeplete targets and removes a pre-determined set of unwanted transcripts prior to sequencing. Unlike other depletion technologies AnyDeplete works after library creation, thus it does not deplete the starting sample at all and can be used on very low concentration samples, all the way down to 10pg concentration. By waiting until after library creation to deplete the unwanted transcripts, AnyDeplete also does not introduce any bias into the final library.

In Step 1, whole transcriptome RNA-Seq libraries contain abundant sequences, such as those from rRNA (shown in red), which provide no useful information but create a high background.

In Step 2, after library construction, probes that target unwanted sequences (AnyDeplete probes) are mixed into the sample. The probes are customizable to any abundant transcript with minimal off target effect.

In Step 3, the AnyDeplete probes facilitate degradation of the unwanted sequences.

In Step 4, the remaining informative transcripts are amplified to create the final sequence ready library now highly enriched for sequences of interest and with the background (non-informative, high abundance transcripts) almost completely removed.

Efficient depletion of unwanted transcripts with AnyDeplete

The table shows the results of AnyDeplete on test libraries generated with 100ng of UHR or K562 human total RNA.

In all cases AnyDeplete was able to efficiently eliminate the unwanted reads from between 37 – 65% of the total library down to less than 1% providing more useful data and lower sequencing costs.

Find this technology in:

RNA-Seq Universal Plus mRNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for mRNA-Seq library preparation for sequencing on Illumina sequencers. This kit features a broad input range, UDI, and simple library quantification with NuQuant.

RNA-Seq Universal Plus Total RNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for total RNA-Seq library preparation for sequencing on Illumina sequencers. This kit features broad input range, UDI adaptors, rRNA depletion with AnyDeplete and simple library quantification with NuQuant.

RNA-Seq Universal RNA-Seq Library Preparation Kit with NuQuant

Robust solution for stranded whole transcriptome RNA-Seq library preparation for any sample type and RNA quality, integrated target depletion after library construction and library quantification.




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Tab 05 / Spet
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SPET – Enrich your library for just the regions of interest

Single Primer Enrichment Technology (SPET) for DNA is a customizable solution for targeted sequencing of specific genomic regions of interest. SPET probes interrogate both strands of the target to enable CNV analysis, robust detection of SNPs and viruses, and re-sequencing.

SPET’s ability to target a large number of specific targets at once makes it a powerful tool for SNP Genotyping, allowing unparalleled sequencing efficiency, leading to rapid scalability and the lowest cost per data point available.

Benefits

SPET provides a rapid, flexible, cost effective solution for targeted amplification and genotyping at any scale, across a wide range of input sample concentrations.

  • Use of single primers, rather than primer pairs, greatly simplifies panel design and allows far higher levels of multiplexing.
  • Flexible customization for any target regions, and able to target anywhere from one to thousands of targets per assay.
  • Simple, fast, and cost-effective genotyping solution, capable of interrogate over 100,000 SNPs in a single assay.

SPET Workflow

A set of targeted probes is created specific to sites of interest. These target probes are mixed with the fragmented genomic DNA and allowed to anneal before being extended. This process creates a pool of amplified fragments all containing the target region of interest. The target probes, all contain generic tails tail sequences that will anneal to a set of PCR primer probes.

Once added to the mix, annealed, and extended through subsequent rounds of amplification, the PCR probes will only anneal to and amplify the fragments created in the initial target amplification round. The result is a pool of PCR fragments, all derived from the product of the initial target amplification, that exponentially increases the presence of target sequence in the final sequence-ready library.

In Step 1, starting genomic DNA is fragmented.

Target probes specific to the region of interest are added in Step 2 and hybridized adjacent to the target region on the DNA fragments.

In Step 3, target probes are extended, creating a pool of target fragments each terminating in a target probe.

PCR probes are added to the mix in Step 4 and the pool is amplified. PCR probes are specific for the tails on the ends of the original target probes and so will only amplify the previously created target fragments from the prior step.

Find this technology in:

Target EnrichmentAllegro Targeted Genotyping V2

Allegro provides a fast, scalable, cost-effective approach to perform targeted genotyping-by-sequencing with the ability to interrogate over 100,000 SNPs in a single assay on a wide variety of organisms using next generation sequencing.




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Tab 06 / UDI
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UDI – High throughput (384 sample) multiplexing and an end to index hopping

Tecan’s Unique Dual Index (UDI) adaptors allow you to take full advantage of the increased sequencing capacity of Illumina sequencing instruments and avoid the problems of index hopping with up to 384 pre-plated UDI adaptors.

UDI enables higher sequencing throughput at reduced cost by sequencing many more samples on the same sequencing run. With Tecan’s pre-plated 384 UDI you can multiplex up to 384 samples in a single run.

Dual Indexes vs. Unique Dual Indexes

Simple Dual Indexes enable an increased level of multiplexing, while Unique Dual Indexes allow you to detect index hopping as well as increasing multiplexing. With 384 UDI adaptors get the best of both worlds! 384 sample multiplex and an end to index hopping.

Benefits

Many of Tecan's library preparation kits now come standard with up to 384 pre-plated UDI adaptors. 

  • Maximize your sequencing capacity by multiplexing up to 384 samples.
  • Unique UDI adaptors for each sample, eliminating the need for multiple sample pools.
  • A simple solution to detect and address index hopping issues with Illumina sequencers.
  • Single-use pre-plated adaptors reduce the possibility of cross-contamination.
  • Sample indexing during adaptor ligation reducing the possibility of adaptor contamination in the final library.
  • Pre-plated UDI adaptors included in the library prep kit, no need to buy separate sets of adaptors.

UDI Workflow

Tecan’s Unique Dual Indexes allow you to uniquely label up to 384 samples with a P7 and a P5 barcode, which are attached to the library fragments in a simple ligation step.

Ligation of two unique barcodes to each of the library fragments (one at each end) mean that the fragments are identified twice. This dual labeling is key to the detection of index hopping, a sequencing phenomenon that occurs at a low level with Illumina sequencing instruments and is most prevalent on patterned flow cells like those used by the Illumina NovaSeq.

If left unchecked index hopping can lead to mis-assignment of reads to the incorrect sample.

Using Tecan’s Unique Dual Indexes in library prep kits, enable the detection and elimination of mis-assigned reads from the sequence dataset.

UDI_Workflow

Find this technology in:

RNA-Seq Universal Plus mRNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for mRNA-Seq library preparation for sequencing on Illumina sequencers. This kit features a broad input range, UDI, and simple library quantification with NuQuant.

RNA-Seq Universal Plus Total RNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for total RNA-Seq library preparation for sequencing on Illumina sequencers. This kit features broad input range, UDI adaptors, rRNA depletion with AnyDeplete and simple library quantification with NuQuant.

RNA-Seq Universal RNA-Seq Library Preparation Kit with NuQuant

Robust solution for stranded whole transcriptome RNA-Seq library preparation for any sample type and RNA quality, integrated target depletion after library construction and library quantification.




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Tab 07 / Dimer Free
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DimerFree – Eliminate adapter dimers with no additional work

Adapter dimers are pairs of adapters bound together but not attached to a fragment from your sequence library. These dimers are particularly problematic because they can still bind and cluster on the sequencer flow cell, generating useless sequence data and occupying space that a real library fragment might have occupied. Adapter dimers are especially problematic at low input concentrations where there may not be enough library fragments to occupy all of the adapters.

All Tecan library prep kits come with our DimerFree technology included. Effectively removing unwanted adapter dimers and eliminating the problem.

Benefits

DimerFree efficiently removed unwanted adapter dimers without requiring any additional steps or additional hands-on time. Get all of the benefits of dimer free libraries without any extra work. Included in all Tecan library prep kits as standard.

  • Simple, fast workflow - no adapter dilution of normalization required.
  • High fidelity libraries – enables representative libraries even in the presence of high GC content.
  • Log scale reduction in adapter dimers – no additional bead clean-up required.

DimerFree Workflow

DimerFree Workflow

DimerFree works to eliminate adapter dimers immediately after adapter ligation and prior to amplification. Eliminating dimers from the pool of fragmented DNA prior to amplification means that only the adapter bound sample fragments go forward to amplification, and on to make up the final sequence ready library.

50% more reads obtained with DimerFree

The power of DimerFree is clearly demonstrated in a head-to-head comparison of three different library prep kits.

At all concentrations Tecan with DimerFree produces a much cleaner library, this effect is especially noticeable as the concentration of the input DNA drops.

DimerFree_Competitor Comparision_720x314

Find this technology in:

DNA-SeqCelero™ DNA-Seq Library Preparation Kit

Simple DNA-Seq library preparation kit that enables library construction in three steps. NuQuant library quantification technology included providing accurate library molarity in seconds.

DNA-SeqCelero™ EZ DNA-Seq Library Preparation Kit

Are you looking for a robust, time-saving DNA-Seq workflow? Celero EZ DNA-Seq Library preparation kit is a simple 3-step workflow integrated with NuQuant library quantification that provides a flexible solution for high-throughput DNA-Seq.

RNA-Seq Universal Plus mRNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for mRNA-Seq library preparation for sequencing on Illumina sequencers. This kit features a broad input range, UDI, and simple library quantification with NuQuant.

RNA-Seq Universal Plus Total RNA-Seq Library Preparation Kit with NuQuant

Streamlined solution for total RNA-Seq library preparation for sequencing on Illumina sequencers. This kit features broad input range, UDI adaptors, rRNA depletion with AnyDeplete and simple library quantification with NuQuant.

RNA-SeqRevelo™ RNA-Seq High Sensitivity library preparation kit

Revelo RNA-Seq High Sensitivity library preparation solutions combine proprietary technologies to provide a streamlined solution for whole transcriptome RNA-seq from low-input or poor quality samples for rare transcript deletion and unbiased pathogen discovery.

DNA-SeqOvation®
Ultralow System V2

The DNA-Seq Library Preparation Kit offering fast and scalable production of NGS libraries from degraded samples as low as 10 pg.

For research use only. Not for use in diagnostic procedures.
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