Plant & Animal Genomes XI Conference Plant & Animal Genomes XI ConferenceJanuary 11-15, 2003
Town & Country Convention Center
San Diego, CA
Poster: High-throughput Methods
Automation platforms are designed for processing large numbers of liquid samples which presents a challenge when using samples stored on solid-phase such as FTA® and CloneSaver™ cards. Here we demonstrate the use of the Biomek®2000 automated liquid handler (Beckman Coulter) to apply samples to FTA/CloneSaver cards and also to process them for downstream use. These cards offer a simple and reliable method for isolating and storing DNA from a variety of biological samples including blood, buccal cells, plant tissue and bacterial cultures including plasmid DNA. When samples are applied to FTA/CloneSaver cards, the cells are lysed and the DNA binds to the FTA matrix. After the sample spots have dried, the DNA is stabilized and can be stored at room temperature for years or processed immediately for analysis by PCR based methodologies, STR profiling or Rolling Circle Amplification. We have optimized the Biomek 2000 to process samples on FTA/CloneSaver. To do this, we took 2.0 mm diameter discs from sample spots on the cards including human blood, buccal cells, plant tissue and bacteria carrying plasmid. For genomic DNA studies, processing an entire 96-well plate of discs was completed in about one hour. For plasmid DNA, the processing time was less than 30 minutes. The samples were washed several times rapidly and used for PCR based methods of genetic analysis including STR profiling. For plasmid DNA, punches were prepared for downstream analysis by PCR, Rolling Circle Amplification with the TempliPhi™ Kit (Amersham BioSciences) and for transformation of competent bacteria.