A High-Throughput System for the Rapid Extraction of Plant Genomic DNA for Genome Mapping and Marker-Assisted Breeding Studies

摘要

Simple sequence repeats (SSR) or simple sequence length polymorphisms (SSLP) are PCR-based molecular markers that have been widely used in genomic mapping and marker-assisted selection. SSR markers have made it possible to establish a high-density genetic map and evaluate genes of interest via tight association between markers and phenotypes. Large-scale marker-assisted breeding studies in plants have created the need for a high-throughput system for genomic DNA preparation and analysis from thousands of samples in a segregating population. However, the standard methods for purifying DNA from plant tissues can be laborious, time-intensive, and not readily amenable to automation. Thus, an automated system for the rapid extraction and subsequent amplification and analysis of plant genomic DNA has been developed to facilitate high-throughput genome mapping and marker-assisted breeding studies. This system utilizes Sigma's Extract-N-Amp Plant PCR kit, a novel system for the rapid extraction and subsequent amplification of genomic DNA from plant tissues, and the maize SSR primer set. This extraction system eliminates time-consuming steps such as organic extractions and mechanical disruption, releasing sufficient genomic DNA from plant tissues for direct use in SSR marker analysis.