Long and Accurate: How HiFi Sequencing is Transforming Genomics

• Published in: Genomics, proteomics & bioinformatics Vol. 23; no. 1
• Main Authors: Wang (王博), Bo, Jia (贾鹏), Peng, Gao (高胜寒), Shenghan, Zhao (赵焕焕), Huanhuan, Zheng (郑高洋), Gaoyang, Xu (许林峰), Linfeng, Ye (叶凯), Kai
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 10.05.2025
• Subjects: Animals; Epigenomics; Genomics - methods; High-Throughput Nucleotide Sequencing - methods; Humans; Review; Sequence Analysis, DNA - methods; Long-read sequencing; Genome assembly; Centromere; Complex genomic region; Variant detection
• ISSN: 1672-0229; 2210-3244
• DOI: 10.1093/gpbjnl/qzaf003

Recent developments in PacBio high-fidelity (HiFi) sequencing technologies have transformed genomic research, with circular consensus sequencing now achieving 99.9% accuracy for long (up to 25 kb) single-molecule reads. This method circumvents biases intrinsic to amplification-based approaches, enabling thorough analysis of complex genomic regions [including tandem repeats, segmental duplications, ribosomal DNA (rDNA) arrays, and centromeres] as well as direct detection of base modifications, furnishing both sequence and epigenetic data concurrently. This has streamlined a number of tasks including genome assembly, variant detection, and full-length transcript analysis. This review provides a comprehensive overview of the applications and challenges of HiFi sequencing across various fields, including genomics, transcriptomics, and epigenetics. By delineating the evolving landscape of HiFi sequencing in multi-omics research, we highlight its potential to deepen our understanding of genetic mechanisms and to advance precision medicine.