| 海南普通野生稻ISSR-PCR反应体系的优化 |
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| 引用本文： | 韩飞,侯立恒,李敬,袁潜华.海南普通野生稻ISSR-PCR反应体系的优化[J].贵州科学,2008,26(4). |
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| 作者姓名： | 韩飞 侯立恒 李敬 袁潜华 |
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| 作者单位： | 1. 海南大学热带生物资源教育部重点实验室,海南,海口,570228;海南大学农学院,海南,海口,570228
2. 海南大学农学院,海南,海口,570228 |
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| 基金项目： | 国家863计划项目(2002AA212091) |
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| 摘 要： | 通过不同浓度的Mg2+、模板DNA、dNTPs、Taq酶、引物、退火温度和循环次数的单因素试验,筛选海南普通野生稻最佳ISSR-PCR反应体系为:15μl反应体系中含2.0 mmol/L MgCl2,50 ng模板DNA,200μmol/L dNTPs,1.20 U Taq酶,1.5μmol/L ISSR引物,50 mmol/L KCl 10 mmol/L Tris-HCl(pH8.3).反应程序:94℃预变性5 min;94℃,1 min,52℃,1 min,72℃,2 min,35个循环;72℃延伸10min,4℃保存.
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| 关 键 词： | 海南 普通野生稻 ISSR 反应体系 PCR |
STUDY ON OPTIM IZATION FOR ISSR-PCR REACTION SYS- TEM FOR WILD RICE INHAINAN PROVINCE |
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| HAN Fei,HOU Li-heng,LI Jing-na,YUAN Qian-hua.STUDY ON OPTIM IZATION FOR ISSR-PCR REACTION SYS- TEM FOR WILD RICE INHAINAN PROVINCE[J].Guizhou Science,2008,26(4). |
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| Authors: | HAN Fei HOU Li-heng LI Jing-na YUAN Qian-hua |
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| Affiliation: | 1.Key Laboratory of Tropic Biological Resources with the Ministry of Education;Hainan University;Haikou;570228;2.College of Agriculture;570228 |
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| Abstract: | To optimize ISSR-PCR reaction system for wild rice in Hainan Province,the effects of seven factors Mg2+,dNTPs,Taq DNA polymerse,primer,T-DNA,annealing temperature and cycle were studied at six levels respectively.The results showed that optimum ISSR-PCR reaction system of wild rice was every 15 μL reaction system containing 2.0 mmol/L MgCl2,50ng template DNA,200μmol/L dNTPs,1.20U Taq DNA polymerse,1.5 μmol/L ISSR primer,50 mmol/L KCl,10 mmol/L Tris-HCl(pH 8.3).The above reaction mixture was denatured at 94℃... |
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| Keywords: | Hainan Oryza rufipogon ISSR reaction system PCR |
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