Rain harvesting is becoming more common in the Palestinian Territories as a result of drinking water scarcity. Although it might pose serious human health risk, this water is being consumed without treatment in many areas of the West Bank. The present study evaluates the physicochemical and microbial quality of harvested rainwater that is used as potable water in the West Bank. Samples from roof-harvested rainwater storage tanks (n = 42) were collected in summer (SS) 2006/winter (WS) 2007. Physicochemical parameters measured were: temperature, pH, electrical conductivity, salinity, total dissolved solids, turbidity, nitrate, copper and lead. With few exceptions, all these parameters were within WHO guideline values. All samples (100%) were found to contain coliforms and to be heavily contaminated with heterotrophic bacteria. About 67% of all samples were contaminated with fecal coliforms. Specific PCR technique confirmed the presence of five pathogenic microorganisms that can be ordered according to their prevalence as: Citrobacter (83%) > Acinetobacter (78%) > Aeromonas (52%) > Pseudomonas and Campylobacter (7%). Prevalence of microorganisms in SS was higher than in WS. Although the physicochemical quality of most harvested rainwater samples was in accordance with WHO guidelines for drinking water, stored rainwater was significantly contaminated with bacteria resulting in significant human health risk from infectious diseases.