High capacity thermal cycler for molecular genotyping

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Arabidopsis thaliana plants homozygous for recessive mutant alleles of the hothead (hth) gene can inherit ancestral DNA sequence information that was not present in the chromosomal genome of their parents. This phenomenon is genome-wide and occurs in both coding and non-coding regions of the genome. These findings are completely novel and the mechanism proposed, unprecedented.  This discovery was made possible in large part by advances in molecular DNA technology. With the advent of PCR, the completion of the Arabidopsis genome sequence and advances in oligonucleotide synthesis procedures, it is now trivial to genotype large numbers of individuals rapidly and reliably.  Because these molecular markers are co-dominant it is also possible to sample both chromosomes simultaneous.  Hence, identifying individuals that carry two alleles that are identical or two distinct alleles that differ in sequence by only one nucleotide is very straightforward. In this application funds are requested to purchase a high capacity modular thermal cycler for genotyping populations of mutant hothead plants and their descendents.  I propose to use PCR technology and molecular markers to assay for changes in DNA sequences  in coding as well as  in non-coding regions of the genome. We will use these simple assays to elucidate the salient features of the novel type of genetic instability uncovered in these mutants.  If this process is found in organisms outside of the plant kingdom, it is possible that this research will have significant future medical relevance.