| 槲树DNA SSR-PCR反应体系的正交优化 |
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| 引用本文： | 叶青雷,王玲玲,藏楠,陈丹,王学英.槲树DNA SSR-PCR反应体系的正交优化[J].蚕业科学,2008,34(2):307-311. |
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| 作者姓名： | 叶青雷 王玲玲 藏楠 陈丹 王学英 |
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| 作者单位： | 沈阳农业大学生物科学技术学院,辽宁省农业生物技术重点实验室,沈阳,110161 |
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| 基金项目： | 辽宁省重点实验室专项课题 |
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| 摘 要： | 利用正交设计L16(45)对槲树(Quercus dentata Thunb.)基因组DNASSR-PCR反应体系的5个因素(Tap酶,Mg2+,,模板DNA,dNTP,引物)在4个水平上进行优化试验,筛选出各反应因素的最佳水平,建立了槲树模板DNASSR-PCR反应的最佳体系(20μL):60ng模板DNA,2mmol/LMg2+,0.075U/μLTaq酶,0.4mmol/L dNTP,引物浓度0.1μmol/L。对槲树DNASSR-PCR最佳反应体系的退火温度进行了梯度试验,最佳退火温度为51.3℃。
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| 关 键 词： | 槲树 SSR标记 反应体系 正交设计 |
Optimization of SSR-PCR Reaction System of Quercus dentata by Orthogonal Design |
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| YE Qing-Lei,WANG Ling-Ling,ZANG Nan,CHEN Dan,WANG Xue-Ying.Optimization of SSR-PCR Reaction System of Quercus dentata by Orthogonal Design[J].Acta Sericologica Sinica,2008,34(2):307-311. |
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| Authors: | YE Qing-Lei WANG Ling-Ling ZANG Nan CHEN Dan WANG Xue-Ying |
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| Abstract: | Orthogonal design was used to optimize SSR-PCR amplification system for Quercus dentata Thunb in terms of 5 factors (Taq DNA polymerase, Mg2+, DNA template,dNTP and primer) from 4 levels.An optimal SSR-PCR system for Quercus dentata Thunb was obtained as 60 ng DNA template, 2 mmol/L Mg2+,0.075 U/μL Taq DNA polymerase, 0.4 mmol/L dNTP,and 0.1 μmol/L primer for 20 μL reaction system. The optimal annealing temperature for SSR-PCR reaction system was determined as 51.3 ℃ by gradient PCR. |
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| Keywords: | Quercus dentata Thunb SSR markers Reaction system Orthogonal design |
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