PacBio SMRT Sequencing Technology

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PacBio SMRT Sequencing Technology

As a leading genomics company, CD Genomics provides next-generation sequencing and bioinformatics services to pharmaceutical and biotech companies, as well as academic and government agencies around the world, relying on advanced sequencing instruments and rich project experience. We are offering long-read sequencing services from PacBio and Oxford Nanopore, giving researchers a wide range of cutting-edge sequencing services to suit the particular needs of any project. Here, we offer long-read sequencing services which are based on PacBio's SMRT high-fidelity (HiFi) Sequencing technology, the Sequel II System. This technology can produce highly accurate (> 99.9% average concordance accuracy) long sequences up to 30 kb long, helping to identify biologically important structural variants, RNA splice site isoforms of cDNA, and provide insights into hard-to-sequence regions of the genome, among others.

Introduction to PacBio SMRT Sequencing Technology

-Technical Principle of SMRT Sequencing

Pacific BioSciences' Single Molecular Real-Time (SMRT) Sequencing technology is also based on sequencing at the same time as synthesis. It works by using a DNA polymerase to restrict the replication of DNA to microwells (zero-mode waveguides, ZMW) and adding fluorescent tracer markers to various bases, which emit different colored flashes when the fluorescent-labeled nucleotides synthesize the DNA strand. The detection system records a movie of light pulses that are converted to base calls to create a continuous long read from each ZMW. The average sequencing read length can reach 3,000 to 5,000 bases, and the longest can reach 20,000 bases.

Fig.1 Overview of PacBio SMRT Sequencing technology. Fig.1 Overview of PacBio SMRT Sequencing technology.

-The PacBio Sequel Systems

Sequel IIe System Sequel II System Sequel System
Supported SMRT cell SMRT Cell 8M SMRT Cell 8M SMRT Cell 1M
Average data output* ~100Gb ~100Gb ~15Gb
Number of HiFi reads >99%* accuracy Up to 4,000,000 Up to 4,000,000 Up to 500,000
Sequencing modes for genomic samples Direct access to high-fidelity (HiFi) Long Reads High-fidelity (HiFi) Long Reads and Continuous Long Reads (CLRs) -
Sequencing run time per SMRT cell Up to 30 hrs Up to 30 hrs Up to 20 hrs
Recommended species/genome size Human(3Gb), plant, or animal with more than 3Gb of genome size Plant, or animal with less than 3Gb of genome size
Features of all PacBio Sequel Systems
  • -Automated consumable handling with integrated software
  • -Intuitive run setup and monitoring
  • -Flexible run time
  • -Walk away time up to 4 days
  • -Ideal for projects such as rapidly and cost-effectively generating high-quality whole genome de novo assemblies and full-length microbiome analysis.
  • * Number of HiFi reads is dependent upon the insert size and sample quality.
  • * Data output is dependent upon the insert size and sample quality.

-The PacBio Sequel II and IIe Systems

Sequel II and Sequel IIe both generate up to 8x more sequencing data per SMRT-cell and belong to the third-generation PacBio sequencers. The principles of their single-molecule sequencing chemistry are unchanged and their sequence coverage and sequence errors do not show any detectable sequence-specific bias. These PacBio Sequel sequencers enable HiFi reads, which is changing genomics. PacBio's HiFi Sequencing technology has some outstanding advantages, including,

PacBio SMRT Sequencing Technology

  • Extraordinary accuracy (>99.9%, Q50).
  • Even coverage.
  • Genome completeness.
  • Allele resolution, long-range phasing.
  • Best performance for all variant classes, due to low sequencing-context bias and accurate mapping of reads.

The Sequel System has been recognized for its ability to generate longer reads with greater accuracy and throughput at a lower cost, providing a simple and rapid SMRT sequencing process suitable for infectious disease, microbial, agricultural and complex genetic disease research.

If you are interested in our services, please don't hesitate to contact us for more details!

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References

  1. Rhoads, A., & Au, K. F. (2015). "PacBio sequencing and its applications." Genomics, proteomics & bioinformatics, 13(5), 278-289.
  2. Ardui, S., et al. (2018). "Single molecule real-time (SMRT) sequencing comes of age: applications and utilities for medical diagnostics." Nucleic acids research, 46(5), 2159-2168.
For Research Use Only. Not for use in diagnostic procedures.

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