An efficient and rapid kanamycin screening assay for identifying true transgenics in cotton

Abstract

Genetic engineering is the best suitable approach for the generation of insect resistant cotton in private and public funded laboratories. Neomycin Phosphotransferase-II is the most commonly used selectable marker gene that can catabolize to kanamycin resistant protein which helps in screening and differentiation of resistant and sensitive plants. In current paper, the optimal concentration of kanamycin was determined as 200 mg/L in control plants. Based on this standardized a procedure for screening transformed plants by spraying 200 mg/L at seedling stage. The resistant plants were reconfirmed by PCR using Npt II specific primers. About 51% and 45% of kanamycin resistant plants were showed amplification of Npt II gene at T1 and T2 generation respectively. This method is simple and fast which can be exploited for screening large number of seedlings with low cost in effective manner.