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Figure 1 | Plant Methods

Figure 1

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

Figure 1

Mutant production and identification using the TILLING process. M2 population, 10 seeds originating from the first M1 fruit were ground and ultimately DNA was isolated, the M2 population comprises 8225 lines. M3 population, from the second M1 fruit, 8810 lines were grown and selfed, seeds were harvested for 7030 lines and a seedlot subset (10 seeds) was used for DNA extraction. For both M2 and M3 population, DNA was pooled 4 or 8 fold, depending on the selected screening method: CSCE; After Multiplex PCR amplification with fluorescent labelled primers, samples are directly pooled together and loaded on capillaries filled with CAP polymer. Pools containing a mutation are identified using Applied Maths' HDA peak analyser software. HRM; Following PCR amplification in presence of LC-Green+™, pools are analysed for their product melting temperatures.

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Plant Methods

ISSN: 1746-4811