Previous researchers have made some efforts to implement this idea: Ogiso-Tanaka et al. It is thus possible to avoid such overkill and reduce the costs by scaling down the reaction system of library construction. If, following manufacturers’ standard library protocols, library output per sample is one to two orders higher than the minimum concentration, dilution is usually needed during library normalization. For example, our Illumina HiSeq Χ Ten service provider requires pooled libraries with ≥ 3 nM concentration and ≥ 15 μL volume.

Genome sequencers usually accept a wide range of pooled library concentrations for one lane of a flow cell. For example, it costs ~ 50 USD for an Illumina DNA library and ~ 8 USD/Gbp for a bacterium sample by Berry Genomics, Inc. As the sequencing costs decrease faster than that of the library construction, the latter accounts for an increasing proportion of the total costs of genomics projects, especially for small genomes such as those of microorganisms. The per-sample costs for genome sequencing have decreased dramatically in recent years, further promoting larger-scale genomics experiments. Consequently this technology has made possible the detection of mutations both at single-base-pair resolution and at whole-genome scale, and the identification of key genes regulating complex phenotypes (Morozova et al. Following its commercialization in 2008, next-generation genome sequencing has been applied to most branches of biology and has facilitated rapid advancements by removing numerous technical barriers such as sequencing speed, read length, throughput and especially cost.

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