Evaluation of Different Reaction Systems for HRM Analysis in Apple
College of Landscape and Horticulture, Qingdao Agricultural University, Qindao, 266109, P.R., China
Bioscience Methods, 2012, Vol. 3, No. 1 doi: 10.5376/bm.2012.03.0001
Received: 16 Nov., 2011 Accepted: 26 Dec., 2011 Published: 29 Jan., 2012
© 2012 BioPublisher Publishing Platform
This article was first published in Molecular Plant Breeding in Chinese, and here was authorized to translate and publish the paper in English under the terms of Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Preferred citation for this article:
Bai et al., 2012, Evaluation of Different Reaction Systems for HRM Analysis in Apple, Bioscience Methods, Vol.3, No.1 1-6 (doi: 10.5376/bm.2012.03.0001)
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.
Apple (Malus domestica); HRM; Reaction volume; DNA template concentration; Touchdown PCR