title: GSM7967885: Fervidibacter sacchari, beta-glucan, biol 3; Candidatus Fervidibacter sacchari; RNA-Seq
library_construction_protocol: Total RNA was extracted using the Qiagen RNeasy Mini Kit using the enzymatic lysis procedure and on-column DNase treatment. 100 ng of total RNA was used to deplete rRNA using the QiaSeq FastSelect 5S/16S/23S Kit. Using TruSeq stranded mRNA kit (Illumina), the 300 - 400 bp heat-fragmented RNA was reverse transcribed to create the first strand of cDNA with random hexamers and SuperScript II Reverse Transcriptase (Thermo Fisher Scientific) followed by second strand synthesis. The double stranded cDNA fragments were treated with A-tailing, ligation with JGI's unique dual indexed adapters (IDT) and enriched using 10 cycles of PCR. The prepared libraries were quantified using KAPA Biosystems' next-generation sequencing library qPCR kit and run on a Roche LightCycler 480 real-time PCR instrument.