Spore Formation Gene Expression Analysis

RNAs from WT/EV, sleC mutant/EV, spo0A mutant/EV, spoVAC*/EV, spoVAC*/spoVAC, dpaAB mutant/EV, and dpaAB mutant/dpaAB strains grown for 18 h on 70:30 sporulation medium containing thiamphenicol (5 μg/ml) were extracted for quantitative real-time PCR (qRT-PCR) analyses of the spoVAC, spoVAD, spoVAE, dpaA, and dpaB transcripts. The RNA was extracted using a FastRNA Pro Blue kit (MP Biomedical) and a FastPrep-24 automated homogenizer (MP Biomedical). Contaminating genomic DNA was depleted using three successive DNase treatments, and mRNA enrichment was done using an Ambion MicrobExpress bacterial mRNA enrichment kit (Invitrogen), and samples were reverse transcribed as previously described (40 (link)).