Abstract: Cotton refers to those species of genus Gossypium that bears the spinnable seed coat fibres. The proteins and enzymes are the primary product of the genes and hence are most suitable for determination of genetic purity and polymorphism. The biochemical and molecular markers may complement the efforts to reduce the burden to a greater extent, as these are estimates devoid of environmental factor which a cause of error. Especially in the selection of parents based on biochemical and molecular polymorphism, there is marked improvement in the screens for the quantitative traits and understanding their architecture. The banding pattern of biochemical markers revealed that the maximum number of bands were recorded in esterase analysis (fifteen) followed by protein analysis (twelve) whereas, only five bands each were detected in peroxidase and polyphenol oxidase analysis indicating limited polymorphism. The Relative Mobility (Rm) values were ranged from 0.083 to 0.883 (protein), 0.100 to 0.971 (esterase), 0.033 to 0.283 (peroxidase) and 0.048 to 0.206 (polyphenol oxidase).