PacBio Literature

PacBio HiFi reads sequence full-length RNA isoforms along with single-cell barcode and UMI information, revealing isoform diversity at the single-cell level.

Getting the most out of your long-read whole genome sequencing (WGS) workflow requires accurately characterizing the size distribution of your genomic DNA and SMRTbell® libraries. The Agilent Femto Pulse system is the recommended quality control (QC) technology for PacBio long-read sequencing using the new SMRTbell prep kit 3.0 workflow

In this Application Note, we focus on HiFi sequencing of full-length PCR amplicons spanning the 3.3 kb CYP21A2 and its pseudogene, CYP21A1P. PacBio HiFi reads are long (up to 25 kb) and accurate (99.9%) making them well suited to analyze the complexity of CAH-related pathogenic variants, where gene conversions and chimeric genes are the predominant mutation types.

With PacBio® HiFi sequencing, CYP2D6 variation can be captured using highly accurate long reads on a single platform. Amplicon-based HiFi sequencing fully and directly resolves and phases complex loci like CYP2D6 without assembly or inference, allowing for ancestry agnostic haplotype assignment.

PacBio® HiFi reads are long (up to 25 kb) and accurate (99.9%). Targeted HiFi sequencing of amplicons has the ability to span entire genes, allowing for straightforward haplotype construction, detection of structural variants or copy number variants in addition to SNVs and indels. These attributes make targeted HiFi sequencing well suited to analyze both rare and common variants of all types, thus increasing the positive detection rate of thalassemia-related variants.

Highly accurate long-read RNA sequencing reveals novel mechanisms of RNA dysregulation in cancer

Gene editing and HiFi sequencing provide fast, reliable solutions that can accelerate the breeding process, modify target genes to improve traits, improve plant resistances to biotic and abiotic stresses and increase plant architecture to better resist climate change.

Extract HMW DNA for HiFi sequencing for one-stop sample-to-answer workflow and support

AMPure® PB bead size selection is a simple, quick, and scalable approach to deplete molecules less than 5 kb in a SMRTbell® library. For high-quality DNA, the sequencing performance and application metrics for assembly and variant calling are comparable to gel cassette size-selection methods.

Bacteria, viruses, and fungi all affect agriculturally important plant and animal species. Accurate HiFi sequencing allows researchers to study microbes and microbial communities in high resolution. Understanding how microbes and microbial communities can impact the health of plants and livestock allows scientists to maximize food production and quality and protect environmental ecosystems.

Scale your whole genome sequencing (WGS) and epigenome workflows with PacBio® HiFi reads employing a single technician working one day a week. Sequence 25 human genomes at ≥ 30-fold coverage each month using four Sequel® II systems.*

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.

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

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.