| 椰子ISSR体系优化 |
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| 引用本文： | 潘坤,王文泉,吴翼,唐龙祥.椰子ISSR体系优化[J].中国农学通报,2009,25(4):24-29. |
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| 作者姓名： | 潘坤 王文泉 吴翼 唐龙祥 |
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| 作者单位： | 1. 中国热带农业科学院椰子研究所,海南文昌,571339
2. 中国热带农业科学院热带生物技术研究所,海南海口,571101 |
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| 基金项目： | 农业部南亚办热带作物种质资源保护项目，科技部国家科技基础条件平台重点项目， |
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| 摘 要： | 本实验旨在获得最佳的椰子ISSR－PCR反应体系，以海南本地高种和马来西亚高种为实验材料，采用单因素实验法将反应体系的五个主要因素设定五个梯度，根据每个因素量的变化对扩增结果产生的影响进行了研究；并利用梯度PCR仪得出引物807的最适退火温度为56.0℃；最后确定最佳反应体系为：总体积20μL，Taq聚合酶1.0U、Mg2＋浓度即10×Taq Buffer用量2.5mmol/L、模板DNA用量50ng、dNTPs浓度0.2mmol/L、引物浓度0.8μmol/L。
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| 关 键 词： | 椰子 ISSR 体系优化 |
| 收稿时间： | 2008-11-19 |
| 修稿时间： | 2008-12-23 |
Optimization of ISSR-PCR system in coconut(Cocos nucifera L.) |
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| Pan Kun,Wang Wen-quan,Wu Yi,Tang Long-xiang.Optimization of ISSR-PCR system in coconut(Cocos nucifera L.)[J].Chinese Agricultural Science Bulletin,2009,25(4):24-29. |
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| Authors: | Pan Kun Wang Wen-quan Wu Yi Tang Long-xiang |
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| Institution: | Pan Kun ;Wang Wenquan ;Wu Yi;Tang Longxiang (1 Coconut Research Institute of Chinese Academy of Tropical Agricultural Sciences;Wenchang Hainan 571339;2 Institute of Tropical Bioscience and Biotechnology of Chinese Academy of Tropical Agricultural Sciences;Haikou Hainan 571101) |
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| Abstract: | The coconut ISSR-PCR reaction system was optimized, we distilled the total DNA of two coconut cultivars that were Hainan Tall and Malaysia Tall, and the single factor experiment was designed which included five factors with five different concentrations and then the effect of the variations of these concentrations were studied. Meanwhile, we obtained the annealing temperature of primer 807, the result shows the optimal annealing temperature was 56℃, at last, the optimum reaction system for coconut consists of 0.5~2.0U Taq DNA polymerse, 2.5~3.0mmol/L 1 0×Taq Buffer(25mmol/L Mg2＋), 50ng template DNA, 0.2~0.5mmol/L dNTPs and 0.8~1.6μmol/L primer with total 20μL reaction solution. |
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| Keywords: | ISSR |
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