Eight previously designed polymorphic microsatellite loci, namely, Mnc434a, Mnc65b, Mnc441, MnBp5-l-02b, MnBp5-l-30b, MnBp5-2-02b, MnBp8-4-43a, and MnBp8-4-43b, were used to examine their modes of segregation in Mystus nemurus family study. Randomly selected broodstocks from Terengganu population were used to generate F1 population. DNA was isolated from tissue samples of the parents, whilst the whole one-month-old F2 progeny was used to isolate the DNA. Chi-square analysis was performed to test the significance of the results obtained. It was shown that all the eight microsatellite loci examined segregated close to Mendelian inheritance ratios, which would be useful in generating a genetic linkage map for this species.