Dental caries is a transmittable infectious disease with a central role of bacteria instrumenting the onset of caries formation. Human oral cavity harbours numerous microbes which are not yet been able to culture or investigate. The present study was aimed to characterize the microbial population associated with the dental caries using culture dependent and culture independent approaches. Various biochemical characterization including Biofilm Formation analysis, Antibiotic Assay and 16s rRNA gene amplification were performed to investigate the culture dependent analysis of microbial flora associated with dental caries. Biofilm formation test revealed that the oral bacterial strains formed better biofilms. Also, molecular approach of recombinant DNA technology (Metagenomics) was employed for culture independent analysis of microbial flora associated with dental caries