Actinomycetes are a group of organisms which have characteristics of both bacteria and fungi, hence, they are also called as ‘Actinobacter’ and ‘Ray fungi’. Actinomycetes are known for producing novel secondary metabolites like enzymes, anti-biotics, anticancerous agents and play major role in recycling of organic matter. In this present research study, actinomycetes were isolated from 11 different soil samples from different places from college campus by serially diluting and spread plate technique on SCA media. 22 actinomycete isolates were obtained, which were identified by gram staining and biochemical tests using Bergey’s manual. The secondary metabolites of the isolated actinomycetes were screened for anti-microbial activity against 8 clinical pathogens by perpendicular streak method (primary screening) and agar well diffusion method (secondary screening). 2 actinomycetes showed mild anti-microbial activity against Proteus vulgaris and Staphylococcus aureus. By using lysozyme method, DNA from selected 8 actinomycetes was isolated and on study of effect of lysozyme concentration on the quantity of DNA using 3 isolates and it was found that .5mg/1ml (15mg/10ml) lysozyme concentration yields highest amount of DNA. Also RAPD PCR was performed on isolate number 4 and its DNA was successfully amplified

Keywords : Actinomycetes, Bergey’s manual, anti-microbial activity, DNA, lysozyme, nanodrop, RAPD PCR