Documentation
Welcome to the PacBio Documentation page. The table below allows you to search for and download the latest guides, protocols, product information, and more. For any questions about our documentation, please contact us.
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Asset Type
Workflow Step
Product Line
Overview – Sequel Systems Application Options and Sequencing Recommendations
Sequel II System, Sequel System
Analysis, Binding, Sequencing, Template Preparation
Description +
This document provides high-level application workflow guidance for preparing sample, sequencing on the Sequel or Sequel II System, and analysis.
Procedure & Checklist – No-Amp Targeted Sequencing Utilizing the CRISPR-Cas9 System
Sequel II System, Sequel System
Binding, Sequencing, Template Preparation
Description +
This procedure can be used to sequence targeted genomic regions, without amplification, using a CRISPR-Cas9 system. This protocol is applicable for the Sequel and Sequel II Systems.
Analysis Procedure – No-Amp Data Preparation and Repeat Analysis
Sequel II System, Sequel System
Analysis
Description +
This document describes analysis procedures for No-Amp data preparation for repeat analysis.
Reference Guide – Designing CRISPR-Cas9 RNA Oligonucleotides for the No-Amp Targeted Sequencing Procedure
Sequel II System, Sequel System
Template Preparation
Description +
This document provides technical guidance on how to generate and select CRISPR RNA oligonucleotides that are compatible with the No-Amp targeted sequencing procedure.
Procedure & Checklist – Preparing SMRTbell Libraries using PacBio Barcoded Overhang Adapters for Multiplexing Amplicons
Sequel II System, Sequel System
Template Preparation
Description +
The procedure describes a workflow for quickly constructing SMRTbell libraries from PCR products using PacBio Barcoded Overhang Adapters and SMRTbell Express Template Prep Kit 2.0. In this workflow, a barcode is introduced to each amplicon through ligation with a hairpin adapter containing a 16-bp barcode. Once barcoded, amplicons can be pooled and purified for sequencing.
Procedure & Checklist – Preparing SMRTbell Libraries using PacBio Barcoded Universal Primers for Multiplex SMRT Sequencing
Sequel II System, Sequel System
Template Preparation
Description +
The procedure describes a workflow for quickly constructing SMRTbell libraries for multiplexing PCR products using PacBio Barcoded Universal Primers and SMRTbell Express Template Prep Kit 2.0. This workflow uses a 2-step PCR for generating up to 96-barcoded amplicons with the first PCR step requiring user-supplied internal primers that contain a combination of universal and target-specific sequences. In the second PCR step, barcodes are incorporated by using universal sequences tailed with 16-bp PacBio barcode sequences. Once the PCR products are barcoded, they can be pooled into a single tube for SMRTbell library construction with the SMRTbell Express Template Prep Kit 2.0.
Procedure & Checklist – Preparing Multiplexed Microbial Libraries Using SMRTbell Express Template Prep Kit 2.0
Sequel II System, Sequel System
Template Preparation
Description +
This streamlined procedure describes how to prepare multiplexed SMRTbell libraries of microbial genomes for sequencing on the Sequel System using the SMRTbell Express Template Preparation Kit 2.0. Up to sixteen genomes can be multiplexed on the Sequel System and up to 48 genomes can be multiplexed on the Sequel II System.
Procedure & Checklist – Amplification of Full-Length 16S Gene with Barcoded Primers for Multiplexed SMRTbell Library Preparation and Sequencing
Sequel II System, Sequel System
Template Preparation
Description +
This document contains instructions for amplifying and constructing a SMRTbell library using Express 2.0 for sequencing full-length 16S genes on the Sequel Systems.
Sequel System Release Notes (v8.0)
Sequel System
Sequencing
Description +
Release Notes for the Sequel System v8.0 Instrument Control Software.
SMRT Link User Guide (v8.0)
Sequel II System, Sequel System
Analysis
Description +
This document describes how to use Pacific Biosciences’ SMRT Link software (v8.0). SMRT Link is the web-based end-to-end workflow manager for Sequel Systems.
SMRT Link Software Installation (v8.0)
Sequel II System, Sequel System
Analysis
Description +
This document describes the procedure for installing SMRT Link v8.0 on a customer system. This document is for use by Customer IT.
Procedure & Checklist – Preparing HiFi SMRTbell Libraries using SMRTbell Express Template Prep Kit 2.0
Sequel II System, Sequel System
Template Preparation
Description +
This document describes a method for quickly constructing large insert SMRTbell libraries for generating high-accuracy long reads (HiFi reads) on the Sequel II System using the SMRTbell Express Template Prep Kit 2.0.
Procedure & Checklist – Preparing Single-Cell Iso-Seq Libraries Using SMRTbell Express Template Prep Kit 2.0
Sequel II System, Sequel System
Template Preparation
Description +
This procedure can be used to construct Single-Cell Iso-Seq SMRTbell libraries using the SMRTbell Express Template Prep Kit 2.0.
Procedure & Checklist – Using AMPure PB Beads for Size-Selection
Sequel II System, Sequel System
Template Preparation
Description +
This procedure describes how AMPure PB beads can be used to remove short insert SMRTbell templates from SMRTbell libraries.
Procedure & Checklist – Preparing 10 kb Library Using SMRTbell Express Template Prep Kit 2.0 for Metagenomics Shotgun Sequencing
Sequel II System, Sequel System
Template Preparation
Description +
This procedure can be used to prepare 10 kb libraries using Template Prep Kit 1.0 for generating highly accurate long reads from 1 µg sheared and concentrated DNA for metagenomic shotgun sequencing.