Experiment was conducted to determine the influence of marination length using evaporated roselle extract (Hibiscus sabdariffa Linn) on microbiological aspects of beef se’i. The completly randomized design (CRD) with 4 treatments 4 replicates was applied in this experiment. Those treatments were P0 marination length of 12 hours without roselle extract; P1 marination length 6 hours with 5% roselle extract; P2 marination length 12 hours with 5% roselle extract, and P3 marination length 18 hours with 5% roselle extract. The parameters measured were pH, moisture, water activity and total plate count. Data compilled were analyzed using variance analysis and Duncan’s multiple range test. The result of analysis showed that pH and moisture of beef se’i significantly affected by marination length using evaporated roselle extract (P<0.05) but no significant (P>0.05) on water activity and total plate count. It was concluded that beef se’i produced through the marination process with different lengths of time (6, 12 and 18 hours) using evaporated rosela extract, showed characteristics of decreased pH, increased water content with high water activity and total plate count tended to decrease. The best marination length of this study was 12 hours using rosela extract.