A pseudo-colony PCR method for rapid identification of transgenic sugarcane lines at in vitro stage
Ramon Enrique, Florencia Budeguer and Aldo Noguera
Abstract: Obtaining transgenic sugarcane lines in vitro is a laborious, expensive, and time-consuming process. To minimize these disadvantages, the presence of transgenes must be checked as soon as possible. Here, we describe a method for direct PCR amplification of DNA from sugarcane plantlets, using smaller quantities of vegetal material, low-cost lab reagents, and rapid processing. By using this methodology, we were able to identify 100% of the transgenic lines generated. The outlined protocol enabled us to perform an earlier checking of the transgene insertions on genomes of transformed lines reducing costs and labor at in vitro stages.