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PacBio  |  PacBio  |  2023

Detecting cancer-related RNA dysregulation with long-read sequencing

Full-length, single-cell RNA data provides critical insights into the understanding of cancer transcriptomic features, such as isoforms, fusions, and expressed mutations. However, until recently, cell typing based on gene expression has been challenging with long reads due to insufficient sequencing depth. In a recent preprint, Dondi et al. (2022) used transcript concatenation and highly accurate long-read sequencing (HiFi sequencing) to increase sequencing depth. This increase enabled cell type identification without needing short reads, and the detection of many new isoforms. They also detected fusions, cell-specific and cell type-specific isoform usage, and revealed differential isoform expression in tumor and mesothelial cells. The authors conclude that “future studies with similar or increased long-read throughput will not have to rely on parallel short-read sequencing, thereby saving cost and labor.”

In this webinar Arthur Dondi discusses:

  • Detection of cell type-specific known and novel isoforms
  • Capture and quantification of full-length isoforms, mutations and fusions in the same scRNA-seq dataset
  • Detection of a gene fusion that was misclassified in matched short-read data
PacBio  |  PacBio  |  2023

Descubra el poder de las secuencias largas y de alta precisión

Con este seminario web descubrirá como las secuencias largas y de alta precisión le ayudarán a lograr nuevos descubrimientos en sus investigaciones, y cómo el nuevo sistema Revio está revolucionado el mundo de la secuenciación genómica.
  |  PacBio  |  2023

A new standard in repeat expansion detection & resolution of segmental duplications with HiFi reads

PacBio is changing the game of genomics with scalable and affordable, highly accurate long-reads from the Revio system. PacBio HiFi reads robustly detect the full spectrum of variation, moving beyond the limitations of other genomics technologies. In this workshop, speakers will share how HiFi sequencing can be applied to solve important clinical research problems such as fully resolving genes that lie within segmental duplications like SMN1/2 and detecting full-length repeat expansions.
PacBio  |  PacBio  |  2023

Metagenome assembly and characterization of a pooled human fecal reference

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Scientists from PacBio and Zymo Research will present state-of-the art HiFi metagenomic sequencing solutions that will enable microbiome researchers to make important discoveries that are not only high-impact but more precise, reproducible, and resource-efficient than ever.
Chen, Xiao and Henno, Geoff   |  PacBio  |  2023

Tackling challenging genes and their paralogs involved in recessive disorders

Effective carrier and newborn screening methods are critical for solving problems in reproductive medicine and launching treatment plans for early-onset genetic diseases. Clinical research faces a number of well-known challenges to disentangle the complexity of the underlying disease genetics. In this exciting webinar, PacBio scientists will showcase new HiFi sequencing solutions developed to accelerate high-impact discovery in carrier and newborn screening research, including Paraphase, a state-of-the-art software tool for analyzing SMN1/SMN2 and other genes with stunning accuracy.
  |  PacBio  |  2023

MAS-Seq for Single Cell Bioinformatics Walkthrough

The PacBio MAS-Seq for 10x Single Cell 3’ kit provides a 16-fold throughput increase on PacBio’s sequencing systems, enabling isoform resolution for single cell transcriptomics. In this tutorial, we demonstrate how to use PacBio’s SMRT Link software to analyze MAS-Seq single cell data to produce gene- and isoform-level count matrices that can be used with tertiary analysis tools such as Kana, Seurat, and CellTypist for cell clustering and cell labeling. For more information on PacBio single-cell isoform informatics, visit https://isoseq.how
PacBio  |    |  2022

Onso system by PacBio – a new standard in short-read sequencing

The Onso system is an innovative benchtop short-read DNA sequencing platform with an unprecedented level of accuracy using PacBio sequencing by binding (SBB) technology. Ultra-high Q40+ data quality allows you to break through limits of detection.
PacBio  |  PacBio  |  2022

Revio and Onso – long-read and short-read sequencing with exceptional accuracy

The Revio long-read sequencer introduces long reads at a lower cost and higher throughput. The Onso short-read sequencer delivers short reads with near-perfect accuracy. Combined, the completeness of long reads with the accuracy of short reads provides the most comprehensive view of genomes, transcriptomes, and epigenomes.
Hanscom, Terrence and Selby, Donald and Tseng, Elizabeth  |  PacBio  |  2022

Highly Accurate HiFi Reads for Discovery, Design, and Manufacturing of AAV Gene Therapy

Gene therapy is at an inflection point and experiencing tremendous growth. High accuracy and complete visibility are critical to the success of novel vector discovery, vector design, and manufacturing quality control for gene therapy products. This webinar illustrates why PacBio HiFi sequencing is the ideal solution due to its ability to sequence full length AAV genomes at high accuracy, which allows for detection of fragmentation, mutations, and large structural events. Short reads, qPCR, and less accurate long-read technology can miss important changes that may be present in the AAV sequence, potentially affecting the safety and efficacy of gene therapy products.
Feng, Xiaowen and Portik, Daniel and Wilkinson, Jeremy  |  Dana-Farber Cancer Institute, Harvard Medical School; PacBio  |  2022

HiFi metagenomics: more samples, more species, more MAGs

Learn how to cost-effectively level up your metagenomics research from expert bioinformaticians. PacBio HiFi sequencing is setting a new standard for quality and resolution in metagenome sequencing thanks to significant strides in data analysis tools. New pipelines that leverage HiFi sequencings unique combination of high accuracy and long read length yield low abundance species at a higher rate, with higher resolution and precision, and return more circular, single contig metagenome-assembled genomes (MAGs).
Liu, Christine and Turner, Tychele  |  University of California, San Diego, and Washington University School of Medicine  |  2022

Unlocking the Genome with Long-Read Sequencing in Genetic Disease Research

Follow two scientists on their journey of discovery using whole genome sequencing and single-cell RNA sequencing to shed light on previously unresolved medical mysteries. In this video, they share how they have been able to successfully identify the genetic drivers in disease research by accessing the complete genomic landscape, including regions previously deemed impossible to sequence.
  |  PacBio  |  2022

Introducing a whole new PacBio

To pioneer the future through biology, you have to look beyond the status quo and see the whole picture. At PacBio, we are devoted to empowering your vision, by delivering trustworthy technologies and the highest accuracy data. Our mission is to enable the promise of genomics to better human health. We create the world’s most advanced sequencing systems that provide the insights scientists need to improve our understanding of biology. We proudly support and collaborate with the scientific community as we work together to reveal the full potential of the genome. We strive to miss nothing, so you can do anything.
Tseng, Elizabeth  |  PacBio  |  2021

Allele-Specific, Isoform-Resolution Single-Cell RNA-Seq Analysis Using Long-Read Sequencing on Concatenated Single-Cell Molecules

In this talk, Dr. Elizabeth Tseng demonstrates a throughput increase for the scIso-Seq method by concatenating single-cell molecules, increasing yield a minimum of 6-fold per SMRT Cell 8M. She explains the bioinformatics workflow for analyzing concatenated scIso-Seq data, which begins with de-concatenation, followed by tagging of UMI and barcode information that can be processed by the isoseq3 pipeline for deduplication. Reads are then aligned against the reference genome, followed by SQANTI3 for transcript classification against a reference annotation (ex: GENCODE) which produces an isoform-level sparse matrix to be analyzed with single-cell tools such as Seurat. She also shows how to apply isoform-level phasing using a tool called IsoPhase, by first calling SNPs based on the deduplicated aligned reads, then using the full-length read information to identify the haplotypes, and finally assigning each isoform-haplotype its associated read counts. Since each read can be associated with its distinct cell type, this allows quantifying isoform expressions in an allele-specific and cell type-specific manner. Finally, she shows how this can be visualized using both standard genome browsers like IGV and long read-aware tools such as SWAN.
Kingan, Sarah  |  PacBio  |  2021

HiFiViral: A Novel Method for Surveillance of SARS-CoV-2 that is Robust Across Sample Input Quantities and the Evolution of New Variants

The COVID-19 pandemic continues to be a major global epidemiological challenge with the ongoing emergence of new strain lineages that are more contagious, more virulent, drug-resistant, and in some cases evade vaccine-induced immunity. In response, the HiFiViral SARS-CoV-2 kit was developed as a scalable solution for the Sequel II and Sequel IIe Systems. Unlike amplicon sequencing, the HiFiViral SARS-CoV-2 kit uses tiled probes, resulting in robust genome coverage across varying viral input quantities despite the presence of new variants. The use of highly accurate long reads, or HiFi reads, enables comprehensive variant detection, including single nucleotide variants, indels, and structural variants, as well as phasing of variants if multiple strains are present in samples. The fully kitted solution contains all reagents needed for viral enrichment and barcoding up to 384 samples, which can be pooled in a single SMRTbell library and run on a single SMRT Cell 8M. The add-only workflow requires two days in the lab with overnight sequencing and analysis and may be automated to achieve a turnaround time of 24 hours. SMRT Link analysis includes variant calling, HiFi read depth plots, the detection of multiple strains in samples, and consensus sequences ready for submission to public databases. In the talk, Dr. Sarah Kingan, demonstrates performance across a broad range of sample Ct, the accuracy of our variant calling method for viral controls, including variants of concern, and the ability to detect multiple variants down to 20% minor frequency. HiFiViral for SARS-CoV-2 is a cost-effective, convenient, and accurate method for viral sequencing, well-suited for scalable surveillance of a rapidly evolving virus to inform public health decision making.
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