Efficient high-throughput DNA extraction protocol for marker-assisted selection in radish (Raphanus sativus L.).

The self-incompatibility (SI) has been used for hybrid seed production in Brassica crops. Concerning the SI response, the S-locus glycoprotein (SLG) and the S-locus receptor kinase (SRK) genes were reported to control the SI response in the stigmatic side. Many primers have been developed using sequence of these two specific genes in Brassica crops. The first step of applying these primers into molecular analysis of SI is DNA extraction. In order to analyse thousands of sample in a short time, finding fast and efficient DNA extraction method was required. In this study, four kinds of DNA extraction methods (xanthogenate buffer, FB plate with xanthogenate buffer, Dneasy 96-plant kit, and Wizard Magnetic 96 DNA plant system) were compared for efficiency in radish. All four PCR amplification results show target bands, thus these four methods were considered good for PCR-based selection in radish. Comparing cost, labour, DNA quality and PCR amplification rate, a method using FB plate with xanthogenate buffer was proved efficient for DNA extraction in radish.