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.
2022 posters
A complete, ultra-low input RNA-seq solution for full-length transcriptome analysis and RNA counting
Objective: RNA sequencing (RNA-seq) is a powerful way to investigate transcriptional highs and lows, allelic origins, and isoform preferences in the transcriptome that can underlie key biological states. One current limitation of single-cell RNA-seq methodologies is either the absence of unique molecular identifiers (UMIs), or the inability to maintain the yield, sensitivity, and reproducibility when UMIs are employed.
Methods: To test the yield, sensitivity, and reproducibility, we benchmarked a new SMART-Seq method, SMART-Seq mRNA (with UMIs), against the existing SMART-Seq v4 Ultra Low Input RNA Kit for Sequencing (SSv4), and the Smart-seq 2 homebrew method (SS2). We included our novel library prep method in the testing to determine if a complete, end-to-end solution improved the data outcome. In addition, we tested the performance of this new method on common automation platforms.
Results: Gene count and read distribution across major RNA-seq output components were comparable between SMART-Seq mRNA (with UMIs) and SSv4. However, the new method showed significantly increased sensitivity compared to the SS2 homebrew method. In addition, we demonstrate that SMART-Seq mRNA (with UMIs) can enable RNA counting, and while optimized for low RNA input, is compatible with single-cell RNA-seq analysis. Finally, we show that SMART-Seq mRNA (with UMIs) is compatible with common automation platforms.
Conclusions: Our data demonstrate that SMART-Seq mRNA (with UMIs) leveraging SMART technology with UMIs for cDNA generation and our unique library preparation protocol, combined with our Cogent NGS analysis software (CogentAP), is a complete, robust, and sensitive solution for full-length transcriptome studies. The inclusion of UMIs allowed for RNA counting without compromising data quality, and lead to superior sensitivity compared to homebrew SS2 chemistries.
Efficient and sensitive high-throughput human B-cell receptor repertoire profiling using SMART technology
Objective: B-cell receptor (BCR) repertoire profiling is increasingly used in health and pathogenic contexts with the goal of biomarker discovery. However, current sequencing technologies are limited in their ability to generate data accurately and reproducibly for all BCR isotypes. To overcome these limitations, we have developed a new kit to accurately profile all heavy (A, D, E, G, M) and light-chain (K, L) isotypes—an end-to-end solution, from library preparation to streamlined data analysis. Here we present data on an updated approach for efficient and high-throughput BCR repertoire profiling of human samples.
Methods: Libraries were prepared from human peripheral blood cells (10 ng–1 μg total RNA) or from B-cells (1 ng–100 ng total RNA) using our new human BCR repertoire profiling kit (~2.5 hours hands-on time). Prepared libraries were then analyzed on the Illumina® Miseq® benchtop sequencer using 300-bp paired-end reads.
Results: For each library, >90% of sequencing reads were on- target while the most highly represented clonotype was found to remain consistent among technical duplicates across a range of input amounts. In comparison to the previous version of our BCR-sequencing kit, the new approach enabled a ~4x increase in total clonotype count observed across various RNA inputs. Furthermore, a sensitivity assay demonstrated that B-cell RNA corresponding to a single clonotype could be detected above background levels when spiked into input total RNA at a relative concentration of 0.001%.
Conclusions: Our new human BCR repertoire profiling kit was found to accurately and reproducibly profile B-cell clones and provide information on the diversity of BCR repertoire in human samples.
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) »
SMART-Seq Stranded Kit performance with ovarian cancer cells (ABRF, March 2019) »
A SMARTer solution to stranded single-cell RNA-seq (ABRF, April 2018) »
Total RNA-seq
Strand-specific transcriptome sequencing for challenging samples (ABRF, March 2014) »
High-throughput automation of single-cell mRNA-seq
Targeted sequencing
Immune profiling
High throughput single cell T-cell receptor profiling by SMART technology (AGBT, February 2018) »
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
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
ChIP-seq for low-input DNA
Single-cell DNA-seq
Whole exome sequencing
High-throughput automation of DNA-seq
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