TILLING- Mutation Detection


Development of a TILLING (Targeting Induced Local Lesions In Genomes) is ongoing at the JGL. Essentially seeds are treated with ethylmethanesulphonate (EMS) to generate a population of plants with random point mutations. By using an effective DNA pooling strategy and amplifying with fluorescently labelled regions of a gene of interest, mismatched heteroduplexes are generated between wild type and mutant DNA products. Heteroduplexes are incubated with the plant endonuclease CEL I, (cleaves heteroduplexs) and the resultant products run on a sequencing platform. Subsequent analysis of the individual plant DNA from the pool DNA identifies the plant bearing the mutation. The regions flanked by the chosen primers in the identified plant is then sequenced to confirm the mutation and the M3 seed is then passed to the user. Genomes being tested for TILLING implementation include Lotus japonicus, Arabidopsis, Barley and Medicago. Development of new and innovative technologies for high throughput mutation detection is also one of our priorities enabling us to offer a quick and cost effective TILLING service.

   
Contact :
David Baker for more information at
JICmutationdetection@jicgenomelab.co.uk