2005, pp 197-210


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TILLING (Targeted Induced Local Lesions in Genomes; McCallum et al. 2000) is a reverse genetic tool for the identification of point mutations in genes of interest within EMS mutagenised populations (Till et al. 2003, Wienholds et al. 2003, Smits et al. 2004), facilitating the efficient screening of a considerable range of mutant alleles for their relation to gene function. TILLING employs the mismatch specific endonuclease CELI which enzymatically identifies any single nucleotide polymorphism between PCR products by recognising sites of mismatch and cleaving the DNA. Several TILLING projects have been instigated in a variety of organisms such as Arabidopsis thaliana, Medicago truncatula, pea, soybean, maize, nematode, rat and zebrafish. In the following chapter, an outline of the procedure and methodology is described, and the initial results from the Lotus japonicus TILLING project are presented.