NGS Service—PacBio Sequel v3

The main reason for using next-generation sequencing is the effective reduction in turnaround time with NGS compared to traditional methods. At the same time, next-generation sequencing makes it possible to analyze the transcriptome and genome of a species in a detailed and holistic way. CD Genomics offers a growing range of services, from library construction to sequencing to complete experimental design and support. Our PacBio Sequel v3-based next-generation sequencing technology can help you advance genome engineering innovation.

Name of the Instrument/Technicial Service

PacBio Sequel v3

MANUFACTURER: Pacific Biosciences

Summary

The Sequel works by detecting DNA replication in real time. Fluors are attached to the gamma-phosphate of the nucleotides. A PacBio library with a single polymerase is loaded into each well. When a nucleotide enters the active site, it can be detected by the sequencer. Basepaired nucleotides have a longer dwell time allowing them to be detected.
The Sequel is able to monitor >500,000 incorporations occurring simultaneously on a SMRTcell. Each run creates a "movie" of the incorporation of bases, collecting data on the intensity of each fluorophore in each well over time. On board computation converts this movie into basecall files.
Using an analysis mode called Circular Consensus Sequencing, high fidelity (HiFi) subreads can be assembled when libraries are sequenced end to end in rolling fashion multiple times. In CCS, a 2 kbp amplicon can easily be sequenced over 10 times end to end over the course of a 20 hr movie. A mode called Continuous Long Read Sequencing simply sequences a subread sequenced over the length of the run, either 10 or 20 hours. In CLR mode, rare lengths of 175 kbp have been seen, but in the Center, it is more typical to see read lengths 50-70 kbp: the longest reads are typically outliers since the average subread length is typically <50 kbp.

Applications

1. DE NOVO ASSEMBLY / LARGE SCALE MAPPING
2. AMPLICON / RESEQUENCING
3. WHOLE TRANSCRIPT
4. BACTERIAL BASE MODIFICATION

CD Genomics, as a specialist sequencing company, has a large range of sequencers in our NGS Core, including Illumina MiSeq, NovaSeq, PacBio, and more. If you require this service, please contact us directly.