High-throughput TILLING (Henikoff S. 2001 ). Seeds are collected and mutagenizd with EMS.M1 plants are allowed to grow and self-pollinated to obtain M2 generations. Each M2 derives from a different M1 plant. M2 DNAs are prepared and M3 seeds are cataloged. To improve efficiency, DNA are pooled. Primers are designed and labeled with IR-dye. After PCR, heating and cool are performed to form mismatches which can be detected and cut by CEL1. Then denature, gel electrophoresis, and scanning with DNA analyzer. Once a single-base-pair allelic variation is screened, use bioinformatic methods and check phenotype. Then the gene function also will be deduced.