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Butanol as fuel component has some advantages compared to ethanol. The Study was aimed at isolation of different types of Clostridium species from the soil and screening for their ability to produce bio-butanol. Soil samples were collected from three different areas in keffi. Clostridium species were isolated using Reinforce Clostridial media and identified using standard microbiological methods, mutation was induced in the Clostridium isolates and 16s rRNA identification technique was used to identify the isolates. Quantitative screening of the Clostridium species isolates capacity in bio-butanol was carried out using sugar cane molasses and waste paper substrate. The result of 16s rRNA of the Clostridium isolated from old barracks was C. acetobutylicium the one isolated from Angwan kwara was C. perfringenes and C. difficile was isolated from Angwan Jaba. Screening for bio-butanol producing ability of the clostridium isolates, shows that C. acetobutylicium produce highest bio-butanol before been exposed to UV light to induce mutation with 9.61± 0.94 mg/l using sugar cane molasses and 4.89± 0.19 mg/l using waste paper followed by C. perfringenes 6.24± 0.61 mg/l using molasses and 3.21±0.11 mg/l using waste paper. After exposing to UV light to induce mutation C. acetobutylicium produced 18.03 ± 0.17 mg/l using molasses and 8.63± 0.06 mg/l using waste paper substrate follow by C. perfringenes 14.19 ± 0.11 mg/l and 5.99 ± 0.31 mg/l. From this study it was observed that C. acetobutylicium has the capacity to produce bio-butanol and also inducing mutation will enhance the capacity of Clostridium species in the production of bio-butanol.