Real-time reverse transcriptase (RT) polymerase chain reaction (PCR) requires data normalization using an appropriate reference gene in order to obtain more reliable results with biological significance. We cloned a partial sequence of elongation factor-1-α (EF1α) and ribosomal protein L17 (RPL17) from Monopterus albus. We investigated the suitability of five commonly used reference genes [18S ribosomal RNA (18S), cytoskeletal protein (β-actin), glyceraldehyde phosphate dehydrogenase (GAPDH), EF1α and RPL17] as potential quantitative reference genes for normalizing real-time RT-PCR data generated in gonads of different developmental stages and in other tissues of M. albus. Analysis of the data indicated that 18S, β-actin and GAPDH are not suitable as reference genes because of their levels of variations of expression. EF1α and RPL17 might be suitable as reference genes in the gonads of different developmental stages as well as in other tissues of M. albus.