There is a lack of appropriate methods for preparing bacterial RNA‐seq library with ultra‐low amount of RNA. To address this issue, we developed miniBac‐seq, a strand‐specific method for high‐quality library construction from sub‐nanogram of total RNA, which is 100‐fold lower than the current benchmark kit and dramatically reduces preparation cost ($28 + $15 × samples). We further demonstrated the high sensitivity of miniBac‐s...