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Figure 1 | BMC Plant Biology

Figure 1

From: Applying genotyping (TILLING) and phenotyping analyses to elucidate gene function in a chemically induced sorghum mutant population

Figure 1

Mutant population generation. Dry seeds of sorghum inbred BTx623 are mutagenized with chemical mutagen EMS and germinated to produce M1 plants. M1 plants are self-fertilized to produce the M2 seeds. One fertile M2 plant from each M2 head row was used to produce M3 progenies. Duplicate leaf samples were collected from the same M2 plant for extracting DNA used in TILLING analysis. Systematic phenotyping was deferred to M3 generation to ensure the mutant phenotypes observed is represented in the M3 seed pool. Ten heads for each M3 head row were pooled as M4 seed pools for public release. Once the DNA is extracted from the mutant population, the DNA is normalized and pooled together as eight-fold pools. The targeted gene is amplified using a forward primer with 700 nm dye label and a reverse primer with an 800 nm dye label attached to the 5' ends. The PCR products are analyzed on LI-COR DNA Analyzer with 700 and 800 nm duel channel following standard TILLING protocol [59].

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