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《Molecular Plant Breeding》 2004-05
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Optimization of SRAP-PCR in Hot Pepper (Capsicum annuum L)

Ren Yu1,2 Wang Deyuan2* Zhang Yindong1 Li Ying2 Wang Hengming21 State Key Laboratory of Tropical Crop Biotechnology, CATAS, Haikou, 5711012 Vegetable Institute, Guangdong Academy of Agricultural Sciences, Guangdong Key Biotech Lab for Fruits and Vegetables, Guangzhou, 510640*Corresponding author, wdy168@tom.com  
Sequence-related amplified polymorphism (SRAP) is a molecular marker technology newly developed by Li and Quiros, which have the characteristics of simplicity, reliability, moderate throughput ratio and distributed evenly in genome. It has been widely applied in the fields of genetic map construction, plant comparative genomics, gene mapping and cloning, cDNA fingerprinting, genetic diversity, heterosis prediction in rice, cotton, rapeseed, potato, apple, citrus, cherry, plum, garlic, lettuce, celery and rice blast fungus. In order to adopt SRAP approach in the hot pepper (Capsicum annuum L) germplasm and heterosis studies, optimizing experiments for the SRAP PCR reactions including PCR protocol, concentrations of DNA template and Mg2+ were carried out in this study. The results showed that the most suitable protocol was initially denaturing at 94℃ for 5 min, then 94℃, 1 min, 35℃, 1min, and 72℃,1 min for the first five cycles, then the annealing temperature is raised to 50℃ for another 35 cycles. The optimum concentrations of DNA template and Mg2+ were 15ng DNA/25?滋l and 2.0mmol/L Mg2+ in this SRAP-PCR system. The new established SRAP-PCR system of hot pepper was fully reproducible and good stability.
【CateGory Index】: S641.3
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