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Friday, February 5, 2021

ASHG CoLab: PacBio HiFi reads for comprehensive characterization of genomes and single-cell isoform expression

In this ASHG 2020 CoLab presentation hear Principal Scientists, Aaron Wenger and Elizabeth Tseng share how highly accurate long reads (HiFi reads) provide comprehensive variant detection for both genomes and transcriptomes. Aaron Wenger describes how new improvements in protocols and analysis methods have increased scalability and accuracy of variant calling. As demonstrated in the precisionFDA Truth Challenge V2, HiFi reads (>99% accurate, 15 kb – 20 kb) now outperform short reads for single nucleotide and structural variant calling and match for small indels. This includes calling >30,000 small variants and >10,000 structural variants missed by short reads, many in medically…

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Friday, February 5, 2021

Educational Video: Variant Detection with HiFi Reads – Understanding Results from the precisionFDA Truth Challenge

In this short video, Aaron Wenger, a Principal Scientist at PacBio, explains what highly accurate long reads, or HiFi reads, are and how they help to detect all variant types including single nucleotide, indels, and structural variants. He goes on to recap the precisionFDA Truth Challenge V2 which used Genome in a Bottle (GIAB) benchmarks to evaluate various sequencing technologies. In the 2020 challenge, when ranked for accuracy, PacBio HiFi reads delivered the highest precision and recall in all categories.

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Friday, February 5, 2021

Video Poster: High-throughput HiFi library workflow for human whole genome sequencing on the Sequel II System

Recent advances in sequencing chemistry and software in the Sequel II System enable generating highly accurate long reads that are up to 25 kb in length with >99% accuracy. The high quality HiFi reads are suitable for variant detection of all types, from single nucleotides to structural variants. PacBio offers an end-to-end solution from sample preparation to data analysis. However, library construction is still a bottleneck making it difficult to implement into a high-throughput workflow for sequencing large number of samples. Input DNA requirements, DNA shearing and size-selection/fractionation are the most critical and challenging steps in the current procedure. In…

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Friday, February 5, 2021

ASHG PacBio Workshop: Latest product and application updates

In this ASHG 2020 PacBio Workshop Jonas Korlach, CSO, shares how the new PacBio Sequel IIe System makes highly accurate long-read sequencing easy and affordable so?all scientists can gain comprehensive views of human genomes and transcriptomes. He goes on to provide updates on the applications including human WGS for variant detection, de novo genome assembly, single-cell full-length RNA sequencing, and targeted sequencing using PCR and No-Amp methods.

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Friday, February 5, 2021

Virtual Global Summit: Using the unique capabilities of the Sequel II System for de novo genome assembly and full-length 16S sequencing

In this PacBio Virtual Global Summit 2020 presentation, Alvaro Hernandez of University of Illinois at Urbana-Champaign shares his experience introducing the Sequel II System to the DNA Sequencing facility at his institution. Hernandez also goes over recommendations to ensure high-quality de novo genome assemblies and benefits of full-length 16S sequencing, compared to the sequencing of variable regions. 

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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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