title: Ice-cap. A high-throughput method for capturing plant tissue samples for genotype analysis.
abstract: High-throughput genotype screening is rapidly becoming a standard research tool in the post-genomic era. A major bottleneck currently exists, however, that limits the utility of this approach in the plant sciences. The rate-limiting step in current high-throughput pipelines is that tissue samples from living plants must be collected manually, one plant at a time. In this article I describe a novel method for harvesting tissue samples from living seedlings that eliminates this bottleneck. The method has been named Ice-Cap to reflect the fact that ice is used to capture the tissue samples. The planting of seeds, growth of seedlings, and harvesting of tissue are all performed in a 96-well format. I demonstrate the utility of this system by using tissue harvested by Ice-Cap to genotype a population of Arabidopsis seedlings that is segregating a previously characterized mutation. Because the harvesting of tissue is performed in a nondestructive manner, plants with the desired genotype can be transferred to soil and grown to maturity. I also show that Ice-Cap can be used to analyze genomic DNA from rice (Oryza sativa) seedlings. It is expected that this method will be applicable to high-throughput screening with many different plant species, making it a useful technology for performing marker assisted selection.
authors.affiliation: Department of Horticulture and Genome Center of Wisconsin, University of Wisconsin, Madison, Wisconsin 53706, USA. fpat@biotech.wisc.edu
authors.forename: Patrick
authors.initials: P
authors.lastname: Krysan
mesh_terms: D005821:Genetic Techniques; D005838:Genotype; D007053:Ice; D010944:Plants / Q000235:genetics*
chemical_list: D007053:Ice