Advances in pharmacogenetic analysis technology have accelerated the movement to incorporate pharmacogenetic analysis data into medicine. Therefore, pharmacists will soon have to provide guidance and raise pharmaceutical questions regarding prescriptions based on patient pharmacogenomic information. The objective of this study was to clarify Japanese pharmacists’ awareness of pharmacogenetics. We conducted a postal questionnaire survey among 372 pharmacists belonging to Tsuruha Holdings. Available data were collected from 268 pharmacists (male [n=133], 40 years old [n=170], drugstore pharmacists [n=182]). Of the pharmacists, 19.0% of the population were aware of the Ethical Guidelines for Human Genome/Gene Analysis Research in Japan, 31.0% of the population had heard either or both the terms “pharmacogenomics” and “pharmacogenetics”, and 16.8% of the population were aware that health insurance covered pharmacogenomic tests performed before prescription in Japan. Only 0.4% indicated that they could raise pharmaceutical questions regarding prescriptions based on patients’ pharmacogenomic information, and 61.2% of the population indicated a preference to undertake pharmacogenomic tests to predict the efficacy or adverse reactions of a drug. We found a need for actions to improve the awareness of pharmacists about pharmacogenetics and create an environment in which pharmacists are able to provide appropriate medical service based on pharmacogenomic information.