The mammalian gut is inhabited by a complex and highly diverse population of bacteria. About 100 trillion microbes are present in the human gut, a number ten times more than the total number of cells in an adult human body. These microorganisms play an important role in several fundamental and crucial processes such as immunity, digestion, synthesis of vitamins, and metabolizing bile acids, sterols, and xenobiotics in the host, thereby influencing human health. Identification and manipulation of these metabolic interfaces is therefore critical. Here, we present a set of methods for manipulation and targeting the 16S rRNA based identification of rodent gut microbiota using Sanger's and next-generation sequencing platforms. Novel methods for manipulation of gut microbiota are also presented. In principle, these methods can be easily adapted to most rodent models for successful screening and manipulation of gut microbiome, to generate a better understanding of their role in metabolic disease.