草业科学2012，Vol.29Issue(1)：59-64,6.
钝裂银莲花ISSR—PCR反应体系优化
Optimization of experiment conditions with ISSR-PCR reaction system in Anemone obtusiloba
摘要
Abstract
The genome DNA extracted by the improved method of CTAB from Anemone obtusiloba collected from Hezuo region in Gansu was used as mode to optimize the ISSR reaction conditions by orthogonal experiment and single factor experiment. This study showed that the optimal conditions of ISSR reaction for A. obtusiloba were 20 μL reaction system, and it contained 10× Buffer 2 μL, Mg2＋ 2.5 mmol . L -1 Taq enzyme 2 U, Genome DNA 2 mg. L-1, dNTPs 0.3 mmol- L-1 and ISSR primer 0.4 mmol .L -1. The process of PCR were 94 ℃ for 5 min, 35 cycles （94 ℃ 30 s,51 ℃ 45 s,72 ℃ 90 s）, and 72℃ for 7 min, and the consult based on primer UBC 807.关键词
钝裂银莲花/ISSR/PCR/反应体系/正交优化Key words
Anemone obtusiloba/ISSR-PCR/amplification system/orthogonal optimization分类
农业科技引用本文复制引用
孙涛,刘左军,孙文斌,刘凤梅,李冰..钝裂银莲花ISSR—PCR反应体系优化[J].草业科学,2012,29(1):59-64,6.基金项目
国家自然科学基金资助项目（30960066）兰州理工大学科研发展基金 （）
