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The study aimed at isolating actinomycetes capable of producing antibiotic like agent once sampled from Nagpur city soil. The study involved selective media as starch casein agar, to isolate actinomycetes, then number of morphological, biochemical tests and 16S rRNA gene sequencing used to identify the species level identity of bacteria. Later on, partially purified bioactive agents tested against pathogen to certain antibacterial features in it. The study recorded that Actinomycetes, Streptomyces sp; Amycolatopsis and Saccharothrix sp profoundly available in tested soil with bio-active agent having nature as alcoholic, phenolic or steroidal.