亚热带农业研究2012，Vol.8Issue(1)：51-56,6.
巨竹ISSR反应体系的建立与优化
Establishment and optimization of ISSR amplification system in Gigantochloa levis
摘要
Abstract
Taking genomie DNA extracted from Gigantochloa levis leaves as the template material, the concentrations of PCR compo- nents, annealing temperature and cycles, whicll affected the ISSR amplification, were optimized with the primer UBC810 （primer sequence ： GAG AGA GAG AGA GAG AT）. The results showed that the 20 μL ISSR reaction system included 40 ng template DNA, 0.6 μmol ·L^-1 primer, 1.0 U Taq DNA polymerase, 2.5 mmol·L^-1 Mg^2＋, 0.25 mmol·L^-1 dNTPs,1×Buffer. The optimal PCR amplification process was 5 minutes at 94 ℃ for predenaturation, followed by 40 cycles ,each with 45 seconds at 94 ℃ for denaturation, 30 seconds at 54.5 ℃ for annealing, 90 seconds at 72℃ for extension,finally extension at 72 ℃ for 10 minutes and holding the samples at 4 ℃.关键词
巨竹/ISSR-PCR/体系优化Key words
Gigantochlochloa levis/ISSR - PCR/system optimization分类
农业科技引用本文复制引用
李炎梅,何天友,陈凌艳,陈礼光,荣俊冬,郑郁善..巨竹ISSR反应体系的建立与优化[J].亚热带农业研究,2012,8(1):51-56,6.基金项目
福建省科技厅重点项目 （）
福建省科技重大项目（2011N5002）. （）
