With easier access to genome sequences and genome editing technology over the last few years, transformation technologies are routinely being used now as a research tool for elucidating gene function. In this chapter, we outline a simple A. tumefaciens-mediated transformation method for the diploid brassica species B. oleracea, using a doubled haploid spring line called AG DH1012. This genotype has become our model genotype for training purposes and for most of our routine work, with a transformation efficiency of 25% (no. independent shoots/no. explants). This easy-to-follow protocol has made for a 100% technology transfer success rate to other researchers and laboratories around the globe. In this chapter, we detail our method using 4-day-old cotyledonary explants, with a step-by-step guide and supplemented with an online video to show the explant and subsequent shoot isolation stages. The video helps to remove any ambiguity that written protocols sometimes have. Primary transgenics ready for DNA extraction and early molecular analysis can be generated within 5–7 weeks from explant isolation.
Part of the book: Genetic Transformation in Crops