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However, the sequencing error rate of 1 per 1000 bp (10\u22123) represents a serious hurdle for research areas focusing on rare mutations, such as somatic mosaicism or microbe heterogeneity. By examining the high-fidelity sequencing methods developed in the past decade, we summarized three major factors underlying errors and the corresponding 12 strategies mitigating these errors. We then proposed a novel framework to classify 11 preexisting representative methods according to the corresponding combinatory strategies and identified three trends that emerged during methodological developments. We further extended this analysis to eight\u00a0long-read sequencing methods, emphasizing error reduction strategies. Finally, we suggest two promising future directions that could achieve comparable or even higher accuracy with lower costs in both NGS and long-read sequencing.<\/jats:p>","DOI":"10.1093\/gpbjnl\/qzae024","type":"journal-article","created":{"date-parts":[[2024,3,11]],"date-time":"2024-03-11T22:25:44Z","timestamp":1710195944000},"source":"Crossref","is-referenced-by-count":10,"title":["Chasing Sequencing Perfection: Marching Toward Higher Accuracy and Lower Costs"],"prefix":"10.1093","volume":"22","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9870-4991","authenticated-orcid":false,"given":"Hangxing","family":"Jia","sequence":"first","affiliation":[{"name":"CAS Key Laboratory of Zoological Systematics and Evolution & State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences , Beijing 100101, China"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5422-8698","authenticated-orcid":false,"given":"Shengjun","family":"Tan","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Zoological Systematics and Evolution & State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences , Beijing 100101, China"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3770-2383","authenticated-orcid":false,"given":"Yong E","family":"Zhang","sequence":"additional","affiliation":[{"name":"CAS Key Laboratory of Zoological Systematics and Evolution & State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences , Beijing 100101, China"},{"name":"University of Chinese Academy of Sciences , Beijing 100049, China"},{"name":"CAS Center for Excellence in Animal Evolution and Genetics, Chinese Academy of Sciences , Kunming 650223, China"}]}],"member":"286","published-online":{"date-parts":[[2024,3,11]]},"reference":[{"key":"2024071201401260900_qzae024-B1","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1038\/nbt1486","article-title":"Next-generation DNA sequencing","volume":"26","author":"Shendure","year":"2008","journal-title":"Nat Biotechnol"},{"key":"2024071201401260900_qzae024-B2","doi-asserted-by":"crossref","first-page":"e113862","DOI":"10.1371\/journal.pone.0113862","article-title":"The accuracy, feasibility and challenges of sequencing short tandem repeats using next-generation sequencing platforms","volume":"9","author":"Zavodna","year":"2014","journal-title":"PLoS One"},{"key":"2024071201401260900_qzae024-B3","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1101\/gr.8.3.186","article-title":"Base-calling of automated sequencer traces using phred. 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