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《Molecular Plant Breeding》 2015-09
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Optimization of High Resolution Melting Analysis System for the Regulatory Region Key SNP of Seed Shattering Gene-qSH1 in Rice

Sheng Xiabing;Tan Yanning;Sun Zhizhong;Yu Dong;Liu Ruifen;Yuan Dingyang;Duan Mei-juan;College of Bioscience and Biotechnology,Hu'nan Agricultural University;State Key Laboratory of Hybrid Rice,Hu'nan Hybrid Rice Research Centre;Hu'nan Academy of Agricultural Sciences;  
High Resolution Melting(HRM) analysis is a newly developed method of fast detection of DNA polymorphism, for its more sensitivity, accuracy and efficiency. Seed shattering is an important agronomy trait affecting seeds harvest. The q SH1 gene is an identified major QTLs controlling seed shattering in rice, in the upstream about 12 kb far from which, a SNP named as q SH1(G/T) was proved essential for seed shattering. In order to establish an optimal HRM genotyping detection system, the effects on HRM of a series of factors including PCR products, DNA concentrations, Mg2+concentrations, annealing temperatures and annealing patteren in the process of PCR were analyzed. The results showed that, the PCR product of 68 bp with the primers(q SH1-1)could make melting peak single and the meltingcurve smooth. Moreover, the combination of 5 ng/μL template DNA, 1.5 mmol/L Mg2+, annealing temperature at 60℃ and a touchdown PCR program would be contributive to an ideal result. These results will lay a foundation for subsequent genotyping and genetic improvement of seed shattering.
【Fund】: 转基因生物新品种培育重大专项(2014ZX0801001B-002);; 湖南省自然科学基金杰出青年项目(2015JJ1026);; 国家863计划项目(2014AA10A604)共同资助
【CateGory Index】: S511;Q943.2
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【Citations】
Chinese Journal Full-text Database 4 Hits
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【Co-citations】
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