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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.
-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 |
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-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,
- 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!
Related Services
- Human Genomics with Long-Read Sequencing
- Animal and Plant Genomics with Long-Read Sequencing
- Microbial Genomics with Long-Read Sequencing
- Transcriptomics with Long-Read Sequencing
- Epigenetics and Methylation Analysis Using Long-Read Sequencing
- Whole-Genome Resequencing with Long-Read Sequencing
- Pre-Made Library Long-Read Sequencing Services
- Long-Read Sequencing Data Analysis Services
References
- Rhoads, A., & Au, K. F. (2015). "PacBio sequencing and its applications." Genomics, proteomics & bioinformatics, 13(5), 278-289.
- 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.