The Nobel laureate Frederick Sanger once said “knowledge of sequences could contribute much to our understanding of living matter”. This statement states the power of bioinformatics, which is the amalgamation of biological and informatic sciences. The significance of bioinformatics or insilico analysis has been reinforced during the pandemic crisis, wherein the computational analysis bolstered the fight against COVID-19 disease. In silico analysis can greatly augment the wet-lab efforts from both in-vivo and in-vitro approaches and includes diverse applications like discovery of new antibiotics or drugs, sustainable agricultural methods, exploring the genetic diversity in different environments. Thus, the promising field of bioinformatics can be explored using some basic practical techniques, which have been explained in this manual. This practical manual of Bioinformatics is a compilation of 12 significant experiments, which help in building a basic foundation in understanding the fascinating subject of Bioinformatics. This subject is offered either as a core paper or an optional paper in the curriculum of an undergraduate course in many universities and colleges across India. The experiments have been presented with a clear approach, which is usually followed in a science practical class and stated in a student’s practical file, which includes : Aim, Theory, followed by the Procedure, Results/Observations and Discussion. The procedure includes respective snapshots of each step in the procedure so that students do not get confused what is being talked about. The results of each experiment have also been explained in greater details, with due emphasis given on even a minute piece of information and it is exclusively for undergraduate students.