Nowadays, drug discovery is a long process which includes target identification, validation, lead optimization, and many other major/minor steps. The huge flow of data has necessitated the need for computational support for collection, storage, retrieval, analysis, and correlation of data sets of complex information. At the beginning of the twentieth century, it was cumbersome to elaborate the experimental findings in the form of clinical outcomes, but current research in the field of bioinformatics clearly shows ongoing unification of experimental findings and clinical outcomes. Bioinformatics has made it easier for researchers to overcome various challenges of time-consuming and expensive procedures of evaluation of safety and efficacy of drugs at a much faster and economic way. In the near future, it may be a major game player and trendsetter for personalized medicine, drug discovery, drug standardization, as well as food products. Due to rapidly increasing commercial interest, currently probiotic-based industries are flooding the market with a range of probiotic products under the banner of dietary supplements, natural health products, food supplements, or functional foods. Most of the consumers are attracted toward probiotic formulations due to the rosy picture provided by the media and advertisements about high beneficial claims. These products are not regulated by pharmaceutical regulatory authorities in different countries of origin and are rather regulated as per their intended use. Lack of stipulated quality standard is a major challenge for probiotic industry; hence there would always be a possibility of marketing of ineffective and unsafe products with false claims. Hence it is very important and pertinent to ensure the safety of probiotic formulations available as over-the-counter (OTC) products for ignorant society. At the same time, probiotic industry, being in its initial stages in developing and underdeveloped countries, requires to ensure safe, swift, and successful usage of probiotics. In the absence of harmonized regulatory guidelines, safety, quality, as well as the efficacy of the probiotic strain does not remain a mandate but becomes a choice for the manufacturer. Hence there is an urgent need to screen already marketed probiotic formulations for their safety with respect to specific strains of probiotic. Various conventional methods used by the manufacturers for the identification of probiotic microbes create a blurred image about their status as probiotics. The present manuscript focuses on a bioinformatics-based technique for validation of marketed probiotic formulation using 16s rRNA sequencing and strain-level identification of bacterial species using Ez Texan and laser gene software. This technique gives a clear picture about the safety of the product for human use.