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,realtime sequencing.Despite it...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,realtime 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 during 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.展开更多
基金supported by the Key R&D Program of Shandong Province,China(grant no.ZR202211070163)the National Natural Science Foundation of China(grant nos.32170574 and 32200249)+1 种基金the Natural Science Foundation of Shandong Province(grant nos.ZR2023QC026 and ZR2023QC106)the Young Taishan Scholars Program and Yuandu Scholars Program.
文摘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,realtime 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 during 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.
