The pursuit of complete telomere-to-telomere(T2T)genome assembly in plants,challenged by genomic complexity,has been advanced by Oxford Nanopore Technologies(ONT),which offers ultra-long,real-time sequencing.Despite its promise,sequencing length and gap filling remain significant challenges.This study optimized DNA extraction and library preparation,achieving DNA lengths exceeding 485 kb;average N50 read lengths of 80.57 kb,reaching up to 440 kb;and maximum reads of 5.83 Mb.Importantly,we demonstrated that combining ultra-long sequencing and adaptive sampling can effectively fill gaps dur-ing assembly,evidenced by successfully filling the remaining gaps of a near-complete Arabidopsis genome assembly and resolving the sequence of an unknown telomeric region in watermelon genome.Collectively,our strategies improve the feasibility of complete T2T genomic assemblies across various plant species,enhancing genome-based research in diverse fields.
Dongdong Lu、Caijuan Liu、Wenjun Ji、Ruiyan Xia、Shanshan Li、Yanxia Liu、Naixu Liu、Yongqi Liu、Xing Wang Deng、Bosheng Li
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Shandong Key Laboratory of Precision Molecular Crop Design and Breeding,Peking University Institute of Advanced Agricultural Sciences,Shandong Laboratory of Advanced Agricultural Sciences in Weifang,Shandong 261325,China
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