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Next-generation sequencing

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Scientific posters

To educate and inform the scientific community about our cutting-edge NGS solutions, Takara Bio staff frequently presents scientific posters at major conferences around the world. Included below are scientific posters featuring methods and data for a wide array of our NGS applications.

2024 posters

Scaling up plate-based single-cell profiling: introducing automated STORM-seq

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Background Full-length, single-cell RNA-seq (scRNA-seq) allows for more complete transcriptomic profiling compared to 5' or 3' end-counting methods. However, labor-intensive, non-modular workflows with high reagent costs limit scalability. Advancements in laboratory automation, protocol design, and reaction miniaturization have overcome some of these constraints. In collaboration with the Van Andel Institute, we present Single-cell TOtal RNA Miniaturized sequencing (STORM-seq), which pairs Takara Bio’s SMART® technology with SPT Labtech’s mosquito® HV genomics liquid handling platform. This miniaturized, plate-based scRNA-seq approach is scalable, automated, and offers a solution for single-cell transcriptomic profiling (i.e., coding/noncoding transcripts, isoforms, gene fusions, etc.) at reduced cost and increased cell throughput.

Methods Single viable K562 cells, peripheral blood mononuclear cells (PMBCs), or renal cell carcinoma cells (RCCs) were sorted into 384-well plates using fluorescence-activated cell sorting (FACS). Following lysis and RNA fragmentation, RNA was reverse transcribed into cDNA. Unique molecular identifiers (UMIs) and unique dual indices (UDIs) were incorporated into the cDNA library following a modified protocol from Johnson and Rhodes et al. (2022, bioRxiv). Miniaturized (1/6) reactions were facilitated using mosquito HV genomics (SPT Labtech). After indexing, pooling, and cleanup, the cDNA libraries were treated with the ZapR™ ribo-depletion technology to remove rRNA-derived cDNAs. The libraries were amplified and sequenced using an Illumina® NextSeq® 500. Sequencing results were analyzed using Cogent™ NGS Software (Takara Bio).

Enabling long-read mRNA-seq for samples with limited input amounts

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Abstract Due to its role in encoding proteins, the ultimate effectors of cell function, and the simplicity of its identification through sequencing, mRNA has become a key indicator in scientific fields such as cancer research, developmental biology, neurobiology, and immunology. Significantly, changes in mRNAs that effect protein function—for example, alternative splicing, SNVs and gene fusions—can be effective biomarkers in these settings.

Third-generation sequencing technologies, such as Oxford Nanopore Technologies (ONT), provide the opportunity to sequence full-length cDNA without the need for fragmentation and hence provide a more complete picture of isoforms present. However, a current limitation of long-read RNA sequencing is the requirement for high input amounts that can be unachievable for some primary sample types, like RNA isolated from tumor samples or sorted blood cancer cells.

SMART® technology (Switching Mechanism at the 5’ end of RNA Template) has emerged as the best solution for detecting and analyzing RNA from a wide range of samples, especially from low input, due to its sensitivity, simplicity, and ability to capture the entire RNA transcript in a single step. An efficient and comprehensive analysis of the full-length cDNA generated by SMART technology has previously been hampered by short-read sequencing, which fragments the resulting output, making it challenging to obtain a full picture of isoform repertoire.

With the improved productivity and accuracy of the ONT platform, combining it with SMART technology in the SMART-Seq® mRNA Long Read (LR) workflow to capture full-length RNA transcripts provides a more complete picture of isoform-specific changes in critical sample types. It is now possible to interrogate up to 96 samples from as little as 10 pg–100 ng of input RNA, which highlights the new opportunities that this technology provides to the field of biomarker discovery.

Previous years' posters sorted by application

Whole transcriptome analysis

Single-cell mRNA-seq

Pushing the limits of single-cell RNA-seq with SMART-Seq single cell technology (ABRF, March 2020) »

Robust and sensitive detection of gene fusions using high-throughput SMART-Seq chemistry on the ICELL8 cx system (AGBT, Feb 2020; ABRF, March 2020) »

SMART-Seq Stranded Kit performance with ovarian cancer cells (ABRF, March 2019) »

A SMARTer solution to stranded single-cell RNA-seq (ABRF, April 2018) »

SMART-Seq v4 Ultra Low Input RNA Kit for the Fluidigm C1 System: improved chemistry for single cell transcriptome studies (NIH Single Cell Analysis Program Investigators Meeting, March 2016) »

Achieving unparalleled sensitivity and reproducibility in single-cell transcriptomics (ABRF, February 2016) »

An improved cDNA library generation protocol for transcriptome analysis from a single cell (ABRF, March 2014) »

Evaluation of commercially available RNA amplification kits at subnanogram input amounts of total RNA for RNA-seq (ABRF, March 2013) »

Total RNA-seq

SMART ligation-free tools for sequencing coding and non-coding RNA from low input samples (Keystone Symposia, February 2017) »

A complete solution for generating stranded RNA-seq libraries from high-input total RNA (ABRF, March 2015) »

SMART tools for strand-specific transcriptome analysis from mammalian total RNA (Keystone Symposia, March 2015) »

Strand-specific transcriptome sequencing for challenging samples (ABRF, March 2014) »

Development of a modified SMART system for robust transcriptome library preparation from limited quantities of compromised samples (ABRF, March 2013) »

High-throughput automation of single-cell mRNA-seq

Automation and miniaturization of low-input RNA-seq sample prep on the Mantis platform (SLAS, January 2016) »

Targeted sequencing

Immune profiling

Efficient high-throughput sequencing for quantitative immune profiling using unique molecular identifiers (ABRF, March 2020) »

High throughput single cell T-cell receptor profiling by SMART technology (AGBT, February 2018) »

A SMARTer approach to profiling the human T-cell receptor repertoire using the ICELL8 single-cell system (Royal Society of Chemistry, Single-Cell Genomics Meeting; September 2016) »

Single-cell T-cell receptor profiling with SMART technology (HudsonAlpha Immunogenomics Conference, September 2016) »

Profiling human TCR repertoire diversity with SMART technology (CAR-TCR Summit, September 2016) »

A SMARTer approach to profiling the human T-cell receptor repertoire (AAI, May 2016) »

A SMARTer approach to profiling the human T-cell receptor repertoire (ABRF, February 2016) »

Small RNA-seq

An unbiased and highly reproducible method for constructing microRNA NGS libraries for accurate expression profiling (ABRF, April 2018) »

Production of an unbiased, highly reproducible small RNA library for NGS using a novel circularization technology (ABRF, March 2017) »

Ligation-free small RNA sequencing libraries from clinical samples using SMART technology (Liquid Biopsies and Minimally-Invasive Diagnostics; September 2016) »

Targeted RNA-seq

A SMARTer approach to gene fusion identification (Next Generation Dx Summit 2017, August 2017) »

Sensitive capture of full-length transcript information with targeted RNA-seq (ASHG, October 2016) »

DNA sequencing

DNA-seq from cfDNA

ThruPLEX HV: a simplified system for preparation of molecular-tagged NGS libraries from FFPE and cell-free DNA (AMP, November 2019; AGBT, February 2020; ABRF, March 2020) »

Confident detection of low-frequency mutations in cell-free DNA using ThruPLEX technology with unique molecular tags (ABRF, April 2018) »

NGS libraries from cell-free DNA containing molecular tags prepared with ThruPLEX technology improve ability to detect rare alleles (ABRF, March 2017) »

ChIP-seq for low-input DNA

Harnessing template-switching technology for preparation of low-input ligation-free sequencing libraries (ABRF, March 2015) »

Harnessing template-switching technology for ChIP-seq applications (CSHL Epigenetics & Chromatin, September 2014) »

Single-cell DNA-seq

Next-generation whole genome amplification methods for CNV and SNV detection from single cells (Molecular Med Tri-Con, March 2019) »

PicoPLEX Gold: a new generation of single cell NGS library with high reproducibility, and greatly improved coverage and fidelity for precision medicine (AMP, October 2018) »

Sequencing single human cells and bacterial DNA using PicoPLEX DNA-seq at low coverage for aneuploidy, CNV, and genotyping applications (AGBT, February 2014) »

Whole exome sequencing

ThruPLEX as a high sensitivity library prep tool for whole exome and target panel sequencing (ASHG, September 2014) »

High-throughput automation of DNA-seq

Utilizing the Rheonix NGS OnePrep Solution to automate the Takara Bio ThruPLEX Tag-Seq HV library preparation kit (ABRF, March 2020) »

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Takara Bio USA, Inc. provides kits, reagents, instruments, and services that help researchers explore questions about gene discovery, regulation, and function. As a member of the Takara Bio Group, Takara Bio USA is part of a company that holds a leadership position in the global market and is committed to improving the human condition through biotechnology. Our mission is to develop high-quality innovative tools and services to accelerate discovery.

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