Development of the lightscanner as a generic platform for novel allele discovery

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The TILLING protocol has been established at Rothamsted Research to identify novel alleles in candidate genes associated with important traits in wheat. The current protocol uses mismatch cleavage of genomic PCR ( Polymerase Chain Reaction) products followed by denaturing polyacrylamide gel electrophoresis. While this is very sensitive and capable of high throughput, the method is expensive, laborious and technically demanding. We recently conceived an alternative method of mutation discovery using high-resolution melt curve analysis to identify point mutations in heteroduplex DNA. We propose to establish this new method on the Lightscanner platform in order to assess the feasibility, utility and throughput of the technique for high throughput mutation detection in wheat. We anticipate that the technique will be transferable to other species and will be particularly useful in diploid organisms where low mutation rates demand high rates of throughput.

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