Whole shotgun metagenomic sequencing for 1,004 individuals belonging to the TwinsUK cohort. Supporting data for: Visconti et al, Interplay between the human gut microbiome and host metabolism

Nextera XT libraries were prepared manually following the manufacturers' protocol (Illumina, PN. 15031942). Briefly, samples were normalized to 0.2 ng/ml DNA material per library using a Quant-iT picogreen assay system (Life Technologies, PN. Q33120) on an AF2200 plate reader (Eppendorf), then fragmented and tagged via tagmentation. Amplification was performed by Veriti 96 well PCR (Applied Biosystems) followed by AMPure XP bead cleanup (Beckman Coulter, PN. A63880). Fragment size for all libraries was measured using a Labchip GX Touch HiSens. Sequencing was performed on an Illumina HiSeq 2500 using SBS kit V4 Chemistry, with a read length of 2x125 bp. Metadata on TwinsUK twin participants are available to bona fide researchers under managed access due to governance and ethical constraints. Raw data should be requested via our website (http://twinsuk.ac.uk/resources-for-researchers/access-our-data/) and requests are reviewed by the TwinsUK Resource Executive Committee (TREC) regularly.

本研究采用手工方式严格遵循制造商提供的实验规程（Illumina，货号PN. 15031942）制备Nextera XT文库。简言之，先通过AF2200微孔板读数仪（Eppendorf）搭配Quant-iT PicoGreen检测系统（Life Technologies，货号PN. Q33120），将每个文库的DNA样品浓度标准化至0.2 ng/ml；随后通过转座酶介导的片段化与标签化反应完成文库构建。扩增步骤采用Veriti 96孔PCR仪（Applied Biosystems）进行，之后使用AMPure XP磁珠完成纯化（Beckman Coulter，货号PN. A63880）。所有文库的片段大小分布均通过Labchip GX Touch HiSens进行检测。测序工作在Illumina HiSeq 2500测序平台上开展，采用SBS V4化学试剂盒，测序读长设置为2×125 bp。由于管理规范与伦理约束，TwinsUK双生子参与者的元数据（metadata）仅对符合资质的研究人员开放受控访问。原始测序数据需通过本研究官方网站（http://twinsuk.ac.uk/resources-for-researchers/access-our-data/）提交申请，申请将由TwinsUK资源执行委员会（TwinsUK Resource Executive Committee，缩写TREC）定期审核。