Metagenomic sequencing of incubated marine sediment
Samples were dual-extracted for both RNA and DNA using the ZymoBIOMICS DNA/RNA Co-extraction Miniprep kit (Zymo Research R2002) following the manufacturers guidelines. Sample nucleic acid concentrations underwent quality control using a Nanodrop 8000 (Thermo Scientific) and a Qubit3 (Invitrogen) with the Broad Range dsDNA Kit (Q33266, Invitrogen). For metagenomic libraries, DNA was first sheared to an average of 350 bp using a Covaris (Covaris) at the Hussman Institute for Human Genomics (HIHG) at the University of Miamis Miller School of Medicine. Sheared DNA was then used in the NEBNext Ultra II DNA Library Prep Kit (New England Biolabs) for Illumina following the manufacturers protocol. Prepared samples were then sent to the HIHG for 150 bp paired-end sequencing on an Illumina NovaSeq with 20 million requested reads per sample.