title: GSM9072993: Cas 7, Replicate 3, 6 Hours; Porphyromonas gingivalis; RNA-Seq
library_construction_protocol: Total RNA was extracted from THP-1 cells co-cultured with either P. gingivalis WT, ∆cas7 for 2h or 6h using a mirVana RNA isolation kit (Life Technologies, New York, USA). Briefly, after removing the media from the wells, cells were incubated with 1 mL of RNAlater (Thermofisher, Pittsburgh, USA) for 5 min. Followed by the addition of mirVana lysis buffer, cells were harvested from the bottom of the wells using a cell scraper and transferred to tubes for cell lysis by bead-beating for 20 seconds three times at maximum speed with diethyl pyrocarbonate (DEPC)-treated zirconia-silica beads (BioSpec Products, Oklahoma, USA). The extraction of RNA from the lysates was conducted as instructed by the manufacturer, following DNAse treatment with Invitrogen DNAse (RNAse free). Illumina's Stranded Total RNA Prep Ligation with Ribo-Zero Plus kit and 10bp unique dual indices (UDI)