RNA sequencing of marine metatranscriptome: Portsmouth Harbor, NH (USA)
Samples were collected and filtered onto 0.2 um PC filters. RNA was extracted from filters using the ZymoBIOMICS DNA/RNA Miniprep kit, which included an on-column DNase I digestion step. DNA was also extracted from these samples. RNA samples were sent to the Michigan State University Research Technology Support Facility Genomics Core for library preparation and sequencing. Total RNA was rRNA depleted using Qiagen FastSelect rRNA depletion kits, followed by cDNA synthesis and library preparation with the KAPA HyperPrep Kit (Roche). Libraries were indexed, pooled, and quantified by Invitrogen Collibri Library Quantification qPCR assays. Sequencing was performed on an AVITI platform (Element Biosciences) to produce 150x150 paired reads.