RNA-seq based transcriptinal profiling of intracellular Brucella abortus and Brucella abortus-infected murine macrophages

We describe the isolation of sufficient Brucella abortus RNA from primary host cell environment using modified reported methods for RNA-seq analysis, and simultaneously characterize the transcriptional profiles of intracellular B. abortus and bone marrow-derived macrophages (BMM) from BALB/c mice at 24 h (replicative phase) post-infection. Each infection and control time point was performed in triplicates using different preparations of BMMs, and processed independently to give three biological replicates. The intracellular B. abortus and infected BMM gene expression were compared to the gene expression of the inoculum and uninfected BMMs, respectively. qRT–PCR validation was performed using SYBR Green assays