Sulfate reducing bacteria (SRB) are obligate anaerobes that can couple their growth to the reduction of sulfate. To systematically provide new insights into SRB biology, we generated a randomly barcoded transposon mutant library in the model SRB Desulfovibrio vulgaris Hildenborough (DvH) and used this genome-wide resource to assay the importance of its genes under a large range of metabolic and stress conditions.