Polymerase chain reaction (PCR) is the most widely used and accepted analytical method for GM detection. Duplex & Multiplex PCR, a derivative of conventional PCR, is reliable, efficient and cost-effective qualitative assay, as fewer reactions are required to test the transgenic nature of a crop by simultaneously detecting the target sequences of the inserted gene construct, i.e. specific transgene, marker genes, promoter and terminator gene sequences in a single PCR assay. The present study reports on the development of qualitative and quantitative PCR assays for detection of commercialized Bt - cotton events, which are being widely cultivated in the North, Central and South zones, i.e. MON531 and MON15985 which are under different stages of field trials in India. Several analytical methods such as methods based on the polymerase chain reaction (PCR) for detecting the transgenic DNA have been developed. DNA has higher stability than proteins, it may also be extracted from processed foods therefore it is preferred analyse for screening of both raw ingredients and processed products. PCR has found real application in GMO detection as an acceptable method for regulatory purposes. Therefore, in the present work attempt has been made to develop and standardize different detection of transgenes Cry1Ac, Cry2Ab, npt-II, 35S Promoter & NOS terminator for Bt - cotton using PCR based strategies and applied to the chosen specific contain any GM genes. Lastly, this effort will be of relevance for timely detection of transgenes in GM foods and products (GMOs) that are still not approved to be imported in India.