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Friday, February 5, 2021

Video Poster: Capture long-read isoform sequencing (Iso-Seq) for uncovering human isoform diversity in the brain and characterizing SARS-CoV2 viral RNAs

Most genes in eukaryotic organisms produce alternative isoforms, broadening the diversity of proteins and non-coding RNAs encoded by the genome. In contrast to other RNA sequencing platforms that rely on short-read sequencing, long accurate reads from PacBio Single Molecule, Real-Time (SMRT) Sequencing can characterize full-length transcripts without the need for assembly and inference. The PacBio isoform sequencing (Iso-Seq) method generates full-length sequences for transcripts up to 10 kb in length, with scalable throughput using barcoding approaches. The Iso-Seq application can be employed for a wide variety of studies, including improvement of gene annotation, identification of novel isoforms and fusion transcripts,…

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Friday, February 5, 2021

Virtual Global Summit: Third generation sequencing for diagnosis of rare disease

In this PacBio Virtual Global Summit 2020 presentation, Neil Miller of Children’s Mercy Hospital in Kansas City describes how third generation sequencing offers sensitive single nucleotide variant detection, variant phasing and the ability to detect large structural variants and complex genomic events which may provide utility in the diagnosis of rare disease. This talk will discuss the use of PacBio HiFi whole genome sequencing in the Genomic Answers for Kids program at Children’s Mercy, Kansas City and how these data are being used to search for disease causing variants in cases that have remained unsolved after clinical exome and whole…

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Friday, February 5, 2021

Virtual Global Summit: Long-range PCR meets long-read sequencing: Tandem technologies for tackling the toughest genes in carrier screening

In this PacBio Virtual Global Summit 2020 presentation, Gary Latham of Asuragen shares how high-prevalence carrier genes associated with disorders such as Fragile X Syndrome, SMA, and Hemophilia include GC-rich repeats, complex structural variants, and/or pseudogenes can derail conventional sequencing methods. Latham reviews these challenges and presents a solution that integrates novel PCR chemistry with SMRT Sequencing to create a unified workflow for carrier screening.

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Friday, February 5, 2021

Video Poster: Improving long-read assembly of microbial genomes and plasmids

Complete, high-quality microbial genomes are very valuable across a broad array of fields, from environmental studies, to human microbiome health, food pathogen surveillance, etc. Long-read sequencing enables accurate resolution of complex microbial genomes and is becoming the new standard. Here we report our novel Microbial Assembly pipeline to facilitate rapid, large-scale analysis of microbial genomes. We sequenced a 48-plex library with one SMRT Cell 8M on the Sequel II System, demultiplexed, then analyzed the data with Microbial Assembly.

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Friday, February 5, 2021

Virtual Global Summit: HIT-scISOseq – High-throughput and high-accuracy single-cell full-length isoform sequencing

In this PacBio Virtual Global Summit 2020 presentation, Tang Chong of BGI shares work on how single-cell isoform sequencing can reveal transcriptomic dynamics in individual cells invisible to bulk- and single-cell RNA analysis based on short-read sequencing. However, current long-read single-cell sequencing technologies have been limited by low throughput and high error rate. Chong introduces HIT-scISOseq for high-throughput single-cell isoform sequencing. This method was made possible by full-length cDNA capture using biotinylated PCR primers, and by a novel library preparation procedure that combines head-to-tail concatemeric full-length cDNAs into a long SMRTbell insert for high-accuracy PacBio Sequencing. HIT-scISOseq represents a high-throughput,…

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Friday, February 5, 2021

Virtual Global Summit: Adventures towards error-free genomes

In this PacBio Virtual Global Summit 2020 presentation, Jennifer Balacco of the Vertebrate Genome Lab at Rockefeller University describes experiences and findings towards pursuing error-free genomes at the Vertebrate Genome Lab. This includes sample prep tips and an example of a VGP high-quality reference genome generated from a museum specimen.

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Friday, February 5, 2021

PagBio Day: New Discoveries in Plant and Animal Sciences

In this PAGBioDay 2021 presentation, Michelle Vierra of PacBio provides the official welcome to PAGBio Day including a presentation on the latest in HiFi sequencing. Michelle is be followed by a keynote presentation by Harris Lewin, Chair of the Earth BioGenome Project at UC Davis, who shares updates and new insights drawn from global conservation initiatives.

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