M220 focused ultra sonicator

2–3 μg genomic DNA per sepsis patient was fragmented on a Covaris M220 focused ultra-sonicator and exomes were enriched by use of Agilent SureSelect XT Human All Exon V5 + UTRs kit, targeting 74,856,280 bp encompassing the coding sequence and untranslated regions of 20,791 human genes. After sequence capture target enrichment, individual libraries were prepared which were quantified and checked for quality by Agilent High Sensitivity DNA chip. Six libraries were pooled each and sequenced on the Illumina HiSeq2500 platform (RapidRun, 2 × 100 bp Paired End). On average, 5.4 × 10⁷ sequence pairs (10.8 Gb) per sample were generated, corresponding to a 215-fold mean depth of coverage per exome (Table S2). A mean of 21% duplicates was detected. DNAs from control individuals were sequenced at the University Kiel after enrichment using the same kit as for the sepsis patients.

Three sediment samples, denoted N1, N4, and N5, were collected from Qinghai Lake, a perennial salt lake located in a structural intermontane depression at the northeastern corner of the Qinghai–Tibetan Plateau (54 (link), 55 (link)), China, in June 2018. Each sample was placed in a sterile plastic bag using a sterile scoop and immediately stored at 4°C. The GPS coordinates of the sampling sites are listed in Data Set S1, Sheet 1. A fraction (10 g by wet weight) of each sample was subjected to DNA extraction as described previously (23 (link), 56 (link)). DNA was purified by using a PowerClean Pro DNA Clean-Up Kit (MO BIO Laboratories), sheared to 400~500 bp in size with the Covaris M220 Focused-Ultrasonicator and quantified by the Agilent 2100 bioanalyzer (Agilent Technologies Inc., USA). The sequencing libraries were constructed using the KAPA Hyper Prep Kit (Kapa Biosystems). Paired-end (PE) sequencing (2 × 250 bp) was conducted on an Illumina Hiseq-2500 platform at Beijing Institute of Genomics, Chinese Academy of Sciences (CAS), Beijing, China. Deep-sea sediments (TVG05 and TVG06), from the Southwest Indian Ocean during the DY125-39 cruise with R/V Dayang No.1, have been previously described by Zheng et al. (23 (link)).