RIb-off rRNA Depletion Kit V2 (Bacteria) is a kit for depleting rRNA from total RNA of Gram-positive and Gram-negative bacteria. This kit is suitable for the total RNA of Gram-positive bacteria and Gram-negative bacteria with the initial template amount of 0.01-1μg. The total RNA sample is processed by rRNA and probe hybridization, RNase H digestion, DNase I digestion and other steps, and finally the rRNA (including 16S and 23S rRNA) is depletd from the total RNA. Retaining mrnas and other non-coding Rnas; The kit is suitable for both intact and partially degraded RNA samples, and the resulting RNA can be used for mRNA and non-coding RNA (e.g., IncRNA) analysis. The optimized reaction system improves the depletional effect and shortens the process time.