| 毛薯ISSR-PCR反应体系的建立 |
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| 引用本文： | 罗欣,周鑫,程文杰,黄小龙,黄东益. 毛薯ISSR-PCR反应体系的建立[J]. 广西农业生物科学, 2010, 0(1): 174-178 |
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| 作者姓名： | 罗欣 周鑫 程文杰 黄小龙 黄东益 |
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| 作者单位： | 海南大学农学院; |
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| 基金项目： | 海南省重点学科建设专项(xkxm0811-03);;高等学校博士学科点专项科研基金(200805650004)共同资助 |
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| 摘 要： | 本研究主要建立毛薯ISSR-PCR的最佳反应体系。研究采用改良CTAB法提取毛薯总基因组DNA,应用单因子实验法设定模板DNA,Mg2＋浓度,dNTP浓度,Tap酶浓度以及退火温度的5个不同梯度,探讨单因素变化对毛薯ISSR-PCR扩增的影响。实验结果表明,毛薯ISSR-PCR最佳反应体系为：总体积20μL,模板DNA为50ng、Taq酶为0.8U、Mg2＋浓度为2.0mmol/L、引物浓度为0.5μmol/L、dNTPs浓度为0.5mmol/L。
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| 关 键 词： | 毛薯 ISSR-PCR 反应体系 |
Establishment of ISSR-PCR System in Dioscorea esculenta (lour) Brukill |
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| Luo Xin Zhou Xin Cheng Wenjie Huang Xiaolong Huang Dongyi Agricultural College of Hainan University,Danzhou. Establishment of ISSR-PCR System in Dioscorea esculenta (lour) Brukill[J]. Journal of Guangxi Agricultural and Biological Science, 2010, 0(1): 174-178 |
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| Authors: | Luo Xin Zhou Xin Cheng Wenjie Huang Xiaolong Huang Dongyi Agricultural College of Hainan University Danzhou |
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| Affiliation: | Luo Xin Zhou Xin Cheng Wenjie Huang Xiaolong Huang Dongyi Agricultural College of Hainan University,Danzhou,571737 |
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| Abstract: | In this study,we have established and optimized ISSR-PCR system of D. esculenta,and we used modified CTAB method to extract its genomic DNA,then setting 5 different gradient by single factor experiment with template DNA,Mg2+ concentration,dNTP concentration,template concentration and annealing temperature,in order to investigate the effect of single-factor change on ISSR-PCR amplification of D. esculenta. The experimental result showed that the optimized ISSR reaction system of D. esculenta was as below,50 ... |
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| Keywords: | Dioscorea esculenta ISSR-PCR Reaction system |
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