GSM4944072: rend_LB_GLB115; Bacillus subtilis; OTHER
5 mL of cell culture was added to 5 mL of cold (-30C) methanol, mixed by inversion and spun down at 3000 rcf for 10 min at 4C. The supernatant was decanted and the cell pellet frozen at -80C. RNA was extracted using the RNAeasy kit (QIAGEN). Ribosomal RNA was depleted using the MICROBExpress kit (Thermo Fisher). The resulting RNA was fragmented for 25 s at 95C using RNA fragmentation reagents (Thermo Fisher, AM 8740). Fragments in the 15 to 45 nt range were selected, dephosphorylated at the 3' end and ligated to a 5' adenylated DNA oligo. After reverse transcription, the single stranded DNA was circularized, and PCR amplified.
GEO Accession
GSM4944072