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This protocol describes a method for isolating and PCR amplifying low-bias long read (8-16 kb) genomic shotgun libraries from small-bodied invertebrates e.g. for de novo genome assembly. It can deliver diverse libraries and contiguous assemblies (100s of kb to several Mb contig N50s are typical) from specimens containing only a few hundred picograms of DNA, such as single meiofaunal specimens (e.g. tardigrades, nematodes, turbellarians). At the same time, it allows users to synthesize and PCR amplify full-length cDNA libraries from the same specimen used for genomic DNA sequencing, permitting evidence-driven genome annotation. The method is compatible with both Oxford Nanopore and PacBio Hifi sequencing, and experienced users will usually be able to complete a batch of specimens within a single week of full-time labwork.
christopher.laumer not provided (Mon,) studied this question.