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Vol 13, Issue 3, 524-530, March 2003

RESOURCES

Large-Scale Discovery of Induced Point Mutations With High-Throughput TILLING

Bradley J. Till1, Steven H. Reynolds1, Elizabeth A. Greene1, Christine A. Codomo1, Linda C. Enns2, Jessica E. Johnson2, Chris Burtner2, Anthony R. Odden1, Kim Young1, Nicholas E. Taylor1, Jorja G. Henikoff1, Luca Comai2 and Steven Henikoff1,3,4

1Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA; 2Department of Biology, University of Washington, Seattle, Washington 98195, USA; 3Howard Hughes Medical Institute

TILLING (Targeting Induced Local Lesions in Genomes) is a general reverse-genetic strategy that provides an allelic series of induced point mutations in genes of interest. High-throughput TILLING allows the rapid and low-cost discovery of induced point mutations in populations of chemically mutagenized individuals. As chemical mutagenesis is widely applicable and mutation detection for TILLING is dependent only on sufficient yield of PCR products, TILLING can be applied to most organisms. We have developed TILLING as a service to the Arabidopsis community known as the Arabidopsis TILLING Project (ATP). Our goal is to rapidly deliver allelic series of ethylmethanesulfonate-induced mutations in target 1-kb loci requested by the international research community. In the first year of public operation, ATP has discovered, sequenced, and delivered >1000 mutations in >100 genes ordered by Arabidopsis researchers. The tools and methodologies described here can be adapted to create similar facilities for other organisms.

4 Corresponding author.

E-MAIL steveh{at}fhcrc.org; FAX (206) 667-5889.

Article and publication are at http://www.genome.org/cgi/doi/10.1101/gr.977903.


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