GSM4060083: PC 1; Bacillus mobilis; RNA-Seq
Cells were scraped from the outermost edges of the growing control or filamentous colonies at 24 h post inoculation directly into RNA Protect Bacterial Reagent (QIAGEN), 1.8 OD 600 was pelleted, and pellets were stored at -80 °C. Pellets were resuspended in 1 ml buffer RLT (QIAGEN) supplemented with 1/100 β-mercaptoethanol and transferred to tubes containing lysing matrix B (MP Biomedicals). Samples were bead bashed in the Bead Ruptor 24 (Omni International) once at speed 7 for 30 s and twice at speed 8 for 30 s, with chilling on ice in between. After a brief spin, the supernatant was mixed with 700 μl 70% EtOH. RNA isolation then proceeded according to the manufacturer's instructions using the RNeasy Mini Kit (QIAGEN). Eluted RNA was further purified with the TURBO DNA-free kit (Thermo Fisher). RNA samples were depleted of rRNA using the RiboZero Bacteria Kit (Illumina). Libraries were prepared using the KAPA Stranded Total RNA Kit (Kapa Biosystems) prior to sending for sequencing. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM4060083