Chasing Sequencing Perfection:Marching Toward Higher Accuracy and Lower Costs

Next-generation sequencing(NGS),represented by Illumina platforms,has been an essential cornerstone of basic and applied research.However,the sequencing error rate of 1 per 1000 bp(10^(−3))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 long-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.