Screening of mutations by tilling in plants

Abstract

TILLING (Targeting Induced Local Lesions IN Genomes) is a well-known reverse genetics technique designed to detect unknown SNPs (single nucleotide polymorphisms) in genes of interest using an enzymatic digestion and is widely employed in plant and animal genomics. The main advantage of this technique is that it allows for the high-throughput identifi cation of an allelic series of mutants with a range of modifi ed functions for a particular gene. In this chapter, we aim to give a detailed introduction of how to establish a TILLING platform for identifying mutants in plants, including generation of a large mutant population, DNA and seed library preparation, mutation identifi cation based on a LI-COR4300 DNA analyzer, and confi rmation of functions of the mutated genes.