A total of 154 clones of reassortants between rotavirus RRV and SA11 were isolated in vitro in order to compare the selection modes of each RNA segment in reassortant formation with those in the in vivo experiment (carried out by Gombold and Ramig) using the same parent strains in which the occurrence of non-random selection of gene segments 3 and 5 was observed. In reassortants isolated from a cross RRV x SA11-L2 (66 isolates) and a cross RRV x SA11-S1 (88 isolates), 36 and 57 different genotypes were identified, respectively. Segregation rates of SA11 RNA segments 1, 5, 8, 9 and 11 were significantly different as compared with those previously reported in the in vivo experiment. While RNA segment 3 was preferentially selected from an SA11 parent, the segregation rate of SA11 gene 5 was relatively low in vitro. Thus, the selection mode of rotavirus RNA segments was found to be different between in vivo and in vitro reassortments.