An optimized toolkit for high-molecular-weight DNA extraction and ultra-long-read nanopore sequencing using glass beads and hexamminecobalt(III) chloride.

ABSTRACT: Since the advent of long-read sequencing, achieving longer read lengths has been a key goal for many users. Ultra-long-read sets (N50 ≥ 100 kb) produced from Oxford Nanopore sequencers have improved genome assemblies in recent years. However, despite progress in extraction protocols and library preparation methods, ultra-long sequencing remains challenging for many sample types. Here, we compare various methods and introduce the FindingNemo protocol that: (1) optimizes ultra-high-molecular-weight (UHMW) DNA extraction and library cleanup by using glass beads and hexamminecobalt(III) chloride (CoHex), (2) can deliver high ultra-long sequencing yield of >20 Gb of reads from a single MinION flow cell or >100 Gb from PromethION devices (R9.4-R10.4 pore variants), and (3) is scalable to using f

SUBMITTER: Cahyani I 

PROVIDER: S-EPMC12047533 | biostudies-literature | 2025 May

REPOSITORIES: biostudies-literature