Fig. 3: Identification of mutant plants using high-resolution melt (HRM) analysis of PCR products derived from pooled plant samples.

a HRM analysis of DNA from pooled samples containing pds-12 with various ratios of mutant (MT) to wild-type (WT) plant tissue. The pooled samples were as follows: a 100% WT plant pool, a 2-plant pool (1MT + 1WT), a 7-plant pool (1MT + 6WT), a 20-plant pool (1MT + 19WT), and a 30-plant pool (1MT + 29WT). Melt curves demonstrate that the 20-plant pool sample can be clearly distinguished from those of the 100% WT pool. b HRM analysis of the 7-plant pool containing different pds mutants. HRM curves of the 100% WT pool (negative control) or 7-plant pools containing a single pds-9, pds-10, pds-11, pds-12, pds-13, or pds-14 mutant demonstrate that HRM analysis is effective to identify single mutants from pooled samples of 7-plants. The WT curve is representative of the three negative control pools used in this single-blind screen. Difference RFU values in b represent the average of three replicates at each temperature point. Error bars show variance for each temperature point for each mutant line