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High resolution melting (HRM) analysis is a newly developed method for fast DNA polymorphism detection. HRM analysis under different reaction volumes, DNA concentrations or polymerase chain reaction (PCR) annealing programs were evaluated for genotyping in apple with cultivar ‘Fuji’ and ‘Telamon’. The result indicated that 5 μL reaction volume was as efficient as 10 μL or 20 μL for revealed the polymorphism of SSR (simple sequence repeat) marker CH03d11, which derived from apple genome, between the two cultivars. Therefore, even DNA concentration as small as 0.25 ng/μL was good enough for PCR amplification and the following HRM detection under a 5 μL reaction volume. Additional study demonstrated that a touchdown PCR program could also performed very well in HRM analysis for polymorphism detection.
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Mudan et al. (2012) studied this question.
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