Grant: Research Tools and Instruments - Category 1 (<$150,000)

My research program is dissecting the molecular mechanisms controlling formation of vein pattern in Arabidopsis thaliana. We identify genes involved in this process by mutation, and then determine the position of the gene within the genome through physical mapping.  Isolation of the gene is accomplished by identifying insertion lines in candidate genes and by sequencing candidate genes from the mutant genome.  Once the gene is isolated, a number of questions about gene expression, gene structure and function and protein localization can be addressed through the creation of transgenes, or through mutating the gene in a targeted fashion and analyzing changes to gene activity.  Each step of this process requires amplification of DNA through the polymerase chain reaction (PCR) using a thermocycler. Our lab (postdoctoral fellow, 2 graduate students, undergraduate students) currently has only one thermocycler - an 8-year old machine with 48 tube capacity.  This machine is very heavily used, and its low capacity limits our throughput on mapping and genotyping experiments. Moreover, if it breaks down, our research stops, because we have no available backup. Therefore, I am requesting 13, 393.70 to purchase an additional thermocycler with 96 tube capacity to increase our throughput and eventually replace our aging machine.