PacBio Literature

Detect, track, and characterize pathogens to a new standard of completeness and precision. PacBio® solutions offer unmatched accuracy and long read lengths to support a wide range of pathogen surveillance applications.

PacBio recommends to use size-selection beads are to remove short DNA fragments and enrich for the long fragments that are most valuable for downstream analysis. Although size-selection beads have many advantages, alternative methods — like gels and DNA precipitation — offer more flexibility in defining a size cutoff. This technical note provides guidance for alternative methods of size selection of WGS libraries prepared with the SMRTbell prep kit 3.0.

HiFi sequencing produces long, accurate reads of the 4 DNA bases — A, C, G, and T — that deliver the most comprehensive characterization of genomes. But HiFi sequencing is not limited to characterizing the genome. It simultaneously measures the epigenome by detecting a fifth base — 5mC at CpG sites — without requiring special library preparation like bisulfite treatment. This detects distinct regional epigenetic patterns, accesses methylation in the full genome, and identifies allele-specific methylation.

PacBio recommends that DNA shearing for whole genome sequencing be performed on the Diagenode Megaruptor 3 system, which is the only system validated by PacBio for shearing, in order to produce consistent HiFi yield and quality. When a Megaruptor 3 system is unavailable, the Covaris g-TUBE presents an alternative shearing method that does not require instrumentation beyond a standard microcentrifuge. This Technical note provides guidelines for using the Covaris g-TUBE for DNA shearing.

Sample preparation is a critical factor for whole genome sequencing (WGS) projects that impacts sequencing yield, read length, and ultimately performance for variant calling and genome assembly. Best practices to optimize performance for PacBio HiFi sequencing are to collect whole human blood, store with potassium EDTA as the anticoagulant for fewer than 2 days at 4°C, and extract DNA from 200 μL of blood using the Nanobind CBB Big DNA kit. When it is not possible to extract from fresh samples, storing samples at 4°C maximizes extraction yield and minimizes degradation for at least 9 days.

The Iso-Seq method utilizes long-read transcript sequencing to reliably capture full-length transcript isoforms without the need for computational assembly. PacBio’s long-read RNA sequencing is also significantly less error-prone than other long-read sequencing technologies, offering more robust and accurate isoform discovery power.9,10 Future research will continue to explore the complex and mostly unexplored landscape of isoforms across cancer types

HiFiViral for SARS-CoV-2 is a simple-to-use, scalable, cost-effective solution for sequencing the entire SARS-CoV-2 genome. This fully kitted solution uses a novel approach that is robust to new variants and comprehensively detects all types of mutations.

With Single Molecule, Real-Time (SMRT) Sequencing and the Sequel Systems, you can affordably assemble reference-quality microbial genomes that are >99.999% (Q50) accurate.

With PacBio Single Molecule, Real-Time (SMRT) Sequencing on the Sequel IIe System you can characterize whole genomes and transcriptomes with just one SMRT Cell. Explore our applications and pricing to get your sequencing project started.

Learn how Single Molecule, Real-Time (SMRT) Sequencing and the Sequel IIe System and will accelerate your research by delivering highly accurate long reads to provide the most comprehensive view of genomes, transcriptomes and epigenomes.

Highly accurate long reads – HiFi reads – with single-molecule resolution make Single Molecule, Real-Time (SMRT) Sequencing ideal for full-length 16S rRNA sequencing, shotgun metagenomic profiling, and metagenome assembly.

This landmark study by members of the Telomere-to-Telomer Consortium is the first fully complete assembly to be produced 20 years after the initial drafts of the human genome.

Learn how PacBio highly accurate long reads enable an improved approach to whole genome sequencing to understand the genetic origins of rare diseases.

With highly accurate long reads (HiFi reads) from the Sequel II or IIe Systems you can comprehensively detect variants in 100s to 1000s of genomes in a year. HiFi reads provide high precision and recall for single nucleotide variants (SNVs), indels, structural variants (SVs), and copy number variants (CNVs), including in difficult-to-map repetitive regions.

PacBio HiFi reads provide both long read lengths (up to 25 kb) and high accuracy (>99.9%) to quickly and affordably generate contiguous, complete, and correct de novo genome assemblies of even the most complex genomes.