Screening of mutations by TILLING in plants.
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 identification of an allelic series of mutants with a range of modified 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 identification based on a LI-COR4300 DNA analyzer, and confirmation of functions of the mutated genes.
Key Laboratory of Plant Germplasm Enhancement and Speciality Agriculture, Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan, China.
Nian
N
Wang
Lei
L
Shi
D004720:Endonucleases / Q000378:metabolism; D005820:Genetic Testing / Q000379:methods*; D018745:Genome, Plant / Q000235:genetics*; D057166:High-Throughput Screening Assays; D016296:Mutagenesis / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D009692:Nucleic Acid Heteroduplexes / Q000235:genetics; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results
D009692:Nucleic Acid Heteroduplexes; D004720:Endonucleases