title: GSM5292362: 3prime3prime-cGAMP-B; Monosiga brevicollis; RNA-Seq
library_construction_protocol: RNA libraries were generated with the Illumina TruSeq® Stranded mRNA Library prep kit (cat # 20020594), using a starting total RNA input of 2-3 µg. To remove contaminating bacterial RNA, samples were first poly-A selected using oligo-dT attached magnetic beads. Following purification, the mRNA was fragmented at 94ºC for 4 minutes, and cleaved RNA fragments were synthesized into cDNA. After an end repair step, UMI adapters (synthesized by IDT) were ligated to the cDNA, and the products were twice purified using AMPure XP beads before amplification.     Library quantity was measured using the Quant-iT™ PicoGreen dsDNA Assay kit by Invitrogen (cat # P7589) and a PerkinElmer Victor X3, 2030 Multilabel Reader. Library quality was verified on an Agilent 2100 Bioanalyzer instrument using Agilent High sensitivity DNA kit (cat # 5067-4626) or DNA 1000 kit (cat # 5067-1504). Libraries were pooled, and sequenced on either the Illumina NovaSeq 6000 system with S4 flowcell and XP PE-100 workflow, or on the Illumina NextSeq 550 system with SE-75 workflow.