Haplotype-resolved genome sequencing promises to unlock a wealth of information in population and medical genetics. However, for the vast majority of genomes sequenced to date, haplotypes have not been determined because of cumbersome haplotyping workflows that require fractions of the genome to be sequenced in a large number of compartments. Here we demonstrate barcode partitioning of long DNA molecules in a single compartment using "on-bead" barcoded tagmentation. The key to the method that we call "contiguity preserving transposition" sequencing on beads (CPTv2-seq) is transposon-mediated transfer of homogenous populations of barcodes from beads to individual long DNA molecules that get fragmented at the same time (tagmentation). These are then processed to sequencing libraries wherein all sequencing reads originating from each long DNA molecule share a common barcode. Single-tube, bulk processing of long DNA molecules with ∼150,000 different barcoded bead types provides a barcode-linked read structure that reveals long-range molecular contiguity. This technology provides a simple, rapid, plate-scalable and automatable route to accurate, haplotype-resolved sequencing, and phasing of structural variants of the genome.

中文翻译：

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在单个试管中使用基于珠子的条形码分配对整个人类基因组进行单倍型定相。

单倍型解析的基因组测序有望解锁人口和医学遗传学领域的大量信息。然而，对于迄今为止已测序的绝大多数基因组，单倍型尚未确定，因为繁琐的单倍型工作流程需要将基因组的一部分在大量的小室中进行测序。在这里，我们演示了使用“珠子”条​​形码标记在单个隔室中对长DNA分子进行条形码分配的方法。珠子上称为“连续性保持转座”测序方法（CPTv2-seq）的方法的关键是转座子介导的条形码均一性从珠子到单个长DNA分子的转移，长条DNA分子同时被片段化（标记）。然后将它们处理到测序文库中，其中源自每个长DNA分子的所有测序读段均共享相同的条形码。具有约150,000种不同条形码条形码类型的长DNA分子的单管批量处理提供了条形码链接的读取结构，该结构揭示了远程分子连续性。这项技术提供了一种简单，快速，可板缩放和自动化的途径，可进行准确的，单倍型解析的测序和基因组结构变异的定相。