| 水稻PCR反应体系的正交设计优化及验证 |
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| 引用本文： | 程芳艳,李春光,刘永巍,孟巧霞,张景龙,宋冬明,刘华招,孟昭河.水稻PCR反应体系的正交设计优化及验证[J].安徽农业科学,2010,38(10):4987-4989. |
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| 作者姓名： | 程芳艳 李春光 刘永巍 孟巧霞 张景龙 宋冬明 刘华招 孟昭河 |
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| 作者单位： | 黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007;黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007;黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007;黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007;黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007;黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007;黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007;黑龙江省农垦科学院水稻研究所,黑龙江,佳木斯,154007 |
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| 基金项目： | 黑龙江省科技厅攻关项目，黑龙江农垦总局项目 |
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| 摘 要： | 目的]筛选经济、稳定性好的水稻PCR反应体系,并检测所选体系在不同的基于PCR反应的分子标记中的通用性。方法]以CTAB法提取的水稻叶片DNA为模板,应用L16（45）正交设计进行PCR反应体系优化。结果]16个不同处理组合均扩增出了清晰谱带,但扩增效果及PCR产量有差异。最经济、适用的体系：20μl反应体系,20ng模板DNA,浓度150μmol/LdNTP,浓度0.2μmol/L引物,1.0UTaqDNA聚合酶,浓度1.5mmol/LMg^2＋,1×Taqbuffer,剩余体积用超纯水补齐。结论]试验确定了水稻PCR优化反应体系,该体系也适用于一些其他基于PCR反应的标记。
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| 关 键 词： | 水稻 PCR体系优化 正交设计 显性标记验证 |
Optimization and Testing of PCR System of Rice by Orthogonal Design |
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| CHENG Fang-yan et al.Optimization and Testing of PCR System of Rice by Orthogonal Design[J].Journal of Anhui Agricultural Sciences,2010,38(10):4987-4989. |
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| Authors: | CHENG Fang-yan |
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| Institution: | CHENG Fang-yan et al (Rice Research Institute,Heilongjiang Academy of L, Reclaimable Sciences,Jiamusi,Heilongjiang 154007) |
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| Abstract: | Objective]The research aimed to screen out the economic and stable PCR system for rice and detect the generality of the selected system in different molecular markers based on PCR.Method]DNA extracted from rice leaves by CTAB method was used as template to optimize PCR system by using L16(45) orthogonal design.Result]Clear bands were amplified from 16 different combinations,but the amplification effects and yield had difference.The most economic and applicable system was as follows:20 ng DNA template,150... |
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| Keywords: | Rice PCR system optimization Orthogonal design Dominant molecular markers testing |
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