| 茄子RAPD分子标记体系的建立与优化 |
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| 引用本文： | 吴雪霞,查丁石,朱宗文,金海军,李贤.茄子RAPD分子标记体系的建立与优化[J].江西农业学报,2011,23(6):51-53. |
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| 作者姓名： | 吴雪霞 查丁石 朱宗文 金海军 李贤 |
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| 作者单位： | 上海市农业科学院园艺研究所、上海市设施园艺技术重点实验室,上海,201106 |
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| 基金项目： | 上海市科技兴农重点攻关项目[沪农科攻字(2009)第2-1号]；现代农业产业技术体系建设专项 |
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| 摘 要： | 采用改良的CTAB法提取茄子基因组DNA,并通过单因素多水平梯度试验,比较筛选RAPD扩增体系的各影响因素,建立了茄子RAPD-PCR的最佳反应体系:20μL反应体系其中含25 mmol/L MgCl2 2.0μL1、0×PCR Buffer 2.0μL、10mmol/L dNTP 0.5μL5、U/μL Taq E 0.2μL0、.1μmol/L Primer 3μL1、0 ng/L模板DNA 3μL、灭菌双蒸水9.3μL。扩增程序为:94℃预变性5 min;94℃变性1 min,37℃退火1 min7,2℃延伸1.5 min4,5个循环;72℃延伸10 min后4℃保存。
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| 关 键 词： | 茄子 RAPD 体系优化 |
Establishment and Optimization of RAPD-PCR Reaction System in Eggplant |
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| WU Xue-xia,ZHA Ding-shi,ZHU Zong-wen,JIN Hai-jun,LI Xian.Establishment and Optimization of RAPD-PCR Reaction System in Eggplant[J].Acta Agriculturae Jiangxi,2011,23(6):51-53. |
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| Authors: | WU Xue-xia ZHA Ding-shi ZHU Zong-wen JIN Hai-jun LI Xian |
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| Institution: | (Horticulture Research Institute,Shanghai Academy of Agricultural Sciences; Shanghai Key Laboratory of Protected Horticultural Technology,Shanghai 201106,China) |
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| Abstract: | The genomic DNA of eggplant（Solanum melongena） was extracted by the modified CTAB method.The factors affecting RAPD results were studied through single factor and multi level tests.An optimal RAPD-PCR reaction system（20 μL） for eggplant was established,and it contained 25 mmol/L MgCl2 2.0 μL,10×PCR Buffer 2.0 μL,10 mmol/L dNTP 0.5 μL,5 U/μL Taq E 0.2 μL,0.1 μmol/L Primer 3 μL,10 ng/L template DNA 3 μL and water 9.3 μL.The appropriate amplification procedure was as follows： pre-denaturing for 5 min at 94 ℃,denaturing for 1 min at 94 ℃,annealing for 1 min at 37 ℃,elongating for 1.5 min at 72 ℃,45 cycles;elongating for 10 min at 72 ℃ and preserving at 4 ℃. |
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| Keywords: | Eggplant RAPD System optimization |
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