Rapid Extraction of PCR-Ready DNA from Plants and Seeds for High Throughput Analysis

The ability to perform rapid, high-throughput plant and seed DNA extractions is highly desirable for plant breeding and other molecular screening applications. We have developed a new method for genomic DNA extraction from plant seeds and leaves, which can be readily adapted to a 96-well extraction format. In less than 10 minutes, PCR-ready DNA can be extracted from up to 10 mg of seed or a 3-5 mm disc of plant leaf material per sample. The simple process consists of adding QuickExtract™ Seed or QuickExtract Plant DNA Solution to the appropriate sample, followed by heating at 65°C for 6 min and 98°C for 2 min. We used the DNA extracted using QuickExtract Solutions as template to amplify single copy genes from a variety of samples for use in both end-point and real-time PCR from fresh or archived plant samples. TILLING (Targeting Induced Local Lesions IN Genomes), SCAR (Sequence Characterized Amplified Region), and other methods for plant mutation and marker discovery are normally dependent on full purification of DNA templates for PCR. In this report, it is shown that the use of rapid DNA extraction of leaf and seed DNA can accelerate and increase throughput of plant genetic studies using amplification of specific loci to obtain DNA for genotyping and other downstream PCR-based assays.