Hydroxyurea (HU) is thought to primarily target ribonucleotide reductase (RNR), therefore inhibiting the conversion of rNTPs into dNTPs and slowing DNA replication. To understand how Bacillus subtilis responds to HU stress, we performed RNA-seq and Tn-seq. Overall design: We obtained transcriptomics following 1 and 2 hours of exposure to hydroxyurea compared to vehicle control from Bacillus subtilis cells.