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Figure 6 | Plant Methods

Figure 6

From: Implementation of two high through-put techniques in a novel application: detecting point mutations in large EMS mutated plant populations

Figure 6

Overview of the mutation screen for ProDH on the M2 population. 8025 EMS mutated families from the M2 population were four-fold flat pooled providing 2112 pools divided among 22 plates of 96 wells. Following PCR, pools were analysed for HRM with a LightScanner® apparatus. HRM analysis identified 87 pools as putatively containing a mutation. Following deconvolution and PCR, the HRM analysis was repeated and 47 single mutant families were sent for Sanger sequencing. Finally based on sequence data 12 mutations were identified. From the first PCR till the identification of the mutants with sequence data, the work load was a total of 5 working days for one person as shown on the left side of the arrows.

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Plant Methods

ISSN: 1746-4811