The PacBio Revio platform supports two sequencing modes: CLR and CCS. Under CCS mode, HiFi reads deliver an average read length of 10–20 kb with a consensus accuracy of no less than 99.9%. Fully compatible with SMRTbell libraries, the Revio system allows four chips to run in parallel. A single instrument run can generate up to 400–480 Gb of high-quality HiFi data. Merging long-read capability with exceptional accuracy, this platform enables robust characterization of structurally complex genomic regions.
The fundamental distinction between CLR and CCS sequencing stems from the number of sequencing passes per DNA template and the implementation of intramolecular consensus correction, resulting in stark differences in read length, base accuracy and applicable research scenarios.
CLR utilizes larger insert sizes with fewer than two full adapter-to-adapter sequencing passes. It achieves average read lengths of 10–30 kb and a subread N50 ranging from 20 to 30 kb. Reads generated via CLR retain raw sequencing error rates, making this mode ideal for large-insert library construction in de novo genome assembly projects.
In contrast, CCS employs shorter insert fragments (typically ~10 kb) with two or more sequencing passes per molecule. Collapsing redundant subreads from the same template greatly elevates base-level accuracy of final CCS reads. This high-fidelity approach is widely applied in genome analysis, full-length transcriptome profiling, full-length 16S sequencing and targeted variant detection studies.
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