Nature Biotechnology
18, 455 - 457 (2000)
doi:10.1038/74542
Targeted screening for induced mutationsClaire M. McCallum1, 2, Luca Comai3, Elizabeth A. Greene1
& Steven Henikoff1, 41
Basic Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109-1024
2
Molecular and Cellular Biology Program, University of Washington, Seattle, WA 98199
3
Department of Botany, University of Washington, Seattle, WA 98199
4
Howard Hughes Medical Institute Research Laboratories, Fred Hutchinson Cancer Research Center, Seattle, WA 98109
Correspondence should be addressed to Steven Henikoff steveh@fhcrc.orgWith the accumulation of large-scale sequence data, emphasis in genomics has shifted from determining gene structure to testing gene function, and this relies on reverse genetic methodology. Here we explore the feasibility of screening for chemically induced mutations in target sequences in Arabidopsis thaliana. Our TILLING (Targeting Induced Local Lesions IN Genomes) method combines the efficiency of ethyl methanesulfonate (EMS)-induced mutagenesis1 with the ability of denaturing high-performance liquid chromatography (DHPLC) to detect base pair changes by heteroduplex analysis2. Importantly, this method generates a wide range of mutant alleles, is fast and automatable, and is applicable to any organism that can be chemically mutagenized.
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