Asset Tag: Amplicon
Quick Reference Card — Loading and Pre-Extension Recommendations for the Sequel II and IIe Systems
Procedure & Checklist — Preparing SMRTbell Libraries using PacBio Barcoded Overhang Adapters for Multiplexing Amplicons
Application brief — HiFi amplicon sequencing
With PacBio® long reads you can easily and cost-effectively sequence full length amplicons that target genes or regions of interest, from several hundred base pairs to kilobase scale. Highly accurate…
High-throughput SMRT Sequencing of clinically relevant targets
Targeted sequencing with Sanger as well as short read based high throughput sequencing methods is standard practice in clinical genetic testing. However, many applications beyond SNP detection have remained somewhat obstructed due to technological challenges. With the advent of long reads and high consensus accuracy, SMRT Sequencing overcomes many of the technical hurdles faced by Sanger and NGS approaches, opening a broad range of untapped clinical sequencing opportunities. Flexible multiplexing options, highly adaptable sample preparation method and newly improved two well-developed analysis methods that generate highly-accurate sequencing results, make SMRT Sequencing an adept method for clinical grade targeted sequencing. The Circular Consensus Sequencing (CCS) analysis pipeline produces QV 30 data from each single intra-molecular multi-pass polymerase read, making it a reliable solution for detecting minor variant alleles with frequencies as low as 1 %. Long Amplicon Analysis (LAA) makes use of insert spanning full-length subreads originating from multiple individual copies of the target to generate highly accurate and phased consensus sequences (>QV50), offering a unique advantage for imputation free allele segregation and haplotype phasing. Here we present workflows and results for a range of SMRT Sequencing clinical applications. Specifically, we illustrate how the flexible multiplexing options, simple sample preparation methods and new developments in data analysis tools offered by PacBio in support of Sequel System 5.1 can come together in a variety of experimental designs to enable applications as diverse as high throughput HLA typing, mitochondrial DNA sequencing and viral vector integrity profiling of recombinant adeno-associated viral genomes (rAAV).
A simple segue from Sanger to high-throughput SMRT Sequencing with a M13 barcoding system
High-throughput NGS methods are increasingly utilized in the clinical genomics market. However, short-read sequencing data continues to remain challenged by mapping inaccuracies in low complexity regions or regions of high homology and may not provide adequate coverage within GC-rich regions of the genome. Thus, the use of Sanger sequencing remains popular in many clinical sequencing labs as the gold standard approach for orthogonal validation of variants and to interrogate missed regions poorly covered by second-generation sequencing. The use of Sanger sequencing can be less than ideal, as it can be costly for high volume assays and projects. Additionally, Sanger sequencing generates read lengths shorter than the region of interest, which limits its ability to accurately phase allelic variants. High-throughput SMRT Sequencing overcomes the challenges of both the first- and second-generation sequencing methods. PacBio’s long read capability allows sequencing of full-length amplicons
Quick Reference Card — Loading and Pre-Extension Recommendations for the Sequel System
User Group Meeting: Lightning Talks – Fast-paced updates from our user base
PacBio Customers present their latest research in short talks at our User Group Meeting. The applications presented span the range of SMRT Sequencing applications from users from around North America.
ASHG PacBio Workshop: Amplicon SMRT Sequencing applications in human genetics
In this ASHG workshop presentation, Stuart Scott of the Icahn School of Medicine at Mount Sinai, presented on using the PacBio system for amplicon sequencing in pharmacogenomics and clinical genomics…
Webinar: Amplicon sequencing with confidence – High-fidelity, long-read PacBio sequencing solutions
In this webinar, Lori Aro and Cheryl Heiner of PacBio describe how high-throughput amplicon sequencing using Single Molecule, Real-Time (SMRT) Sequencing and the Sequel System allows for the easy and…
Webinar: Understanding SARS-CoV-2 and host immune response to COVID-19 with PacBio sequencing
Studying microbial genomics and infectious disease? Learn how the PacBio Sequel II System can help advance your research, with first-hand perspectives from scientists who are investigating SARS-CoV-2 and COVID-19. In…
Webinar: Opportunities for using PacBio Long-read sequencing for COVID-19 research
In this Labroots webinar, Meredith Ashby, Director of Microbial Genomics at PacBio, describes the utility of highly accurate long-read sequencing, known as HiFi sequencing, to understand the SARs-CoV-2 viral genome….
Webinar: Geographic and Temporal Mapping of the SARS-CoV-2 Pandemic in the United States
In this presentation, Michael Levandoski of LabCorp, shares his talk entitled Geographic and Temporal Mapping of the SARS-CoV-2 Pandemic in the United States. Learn more at pacb.com/COVID-19