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 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.

中文翻译：

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纳米孔超长测序和适应性采样刺激植物完整的端粒到端粒基因组组装


提供超长实时测序的 Oxford Nanopore Technologies （ONT） 推进了在植物中追求完整的端粒到端粒 （T2T） 基因组组装，受到基因组复杂性的挑战。尽管前景广阔，但测序长度和间隙填充仍然是重大挑战。本研究优化了 DNA 提取和文库制备，实现了超过 485 kb 的 DNA 长度;平均 N50 读长为 80.57 kb，最高可达 440 kb;最大读取量为 5.83 Mb。重要的是，我们证明超长测序和自适应采样相结合可以有效地填补组装过程中的空白，成功填充近乎完整的拟南芥基因组组装的剩余空白并解析西瓜基因组中未知端粒区域的序列证明了这一点。总的来说，我们的策略提高了跨各种植物物种的完整 T2T 基因组组装的可行性，加强了不同领域基于基因组的研究。