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| 人参基因组RAPD条件的优化 | |
| 引用本文： | 许永华,张国荣,董宇,杜跃中,夏广清,张春颖.人参基因组RAPD条件的优化[J].人参研究,2010,22(1):2-4. |
| 作者姓名： | 许永华 张国荣 董宇 杜跃中 夏广清 张春颖 |
| 作者单位： | 1. 吉林农业大学中药材学院,吉林,长春,130118 2. 吉林人参研究院,吉林,通化,134001 3. 吉林省通化师范学院,吉林通化,134002 |
| 摘 要： | 目的确定药用人参最佳随机扩增多态DNA(RAPD)分析方法。方法以药用人参新鲜叶为材料,研究对影响RAPD反应的各因素进行优化。结果药用人参RAPD反应体系及程序为:在25μL反应体系中,含40ng模板DNA,0.3μmol/L随机引物,0.2mmol/LdNTPs,2.5μL10×PCRBuffer,2.0UTaq酶;扩增程序为:第一步94℃预变性2min,第二步94℃变性1min,第三步36℃退火45s,第四步72℃延伸90s,返回至第二步重复40个循环,第六步72℃延伸10min,最后4℃保存以备电泳。结论建立了人参RAPD的优化反应体系及程序。 |
| 关 键 词： | 人参 DNA提取 RAPD 条件优化 |
DNA extr action from and optimization of RAPD reaction system for ginseng |
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| Authors: | XU Yong-hua Zhang Guo-rong DONG Yu DU Yue-zhong XIA Guang-qing ZHANG Chun-ying |
| Institution: | XU Yong - hua Zhang Guo -rong DONG Yu DU Yue -zhong XIA Guang - qing ZHANG Chun - ying( Jilin Agricultural University, Changchun, Jilin 130118 Jilin Ginseng Institute Tonghua Jilin 134001 Tonghua Normal University Tonghua Jilin 134001 ) |
| Abstract: | Objective To confine the best analysis method by RAPD of the ginseng, the RAPD conditions were discussed Results The system and program of RAPD for ginseng were as follows： The reactions were performed in a volume of 25 μL containing 40ng of DNA, 0. 3 μmol/L primer, 0. 2mmol/LdNTPs, 2. 5 p,L 10 × PCR Buffer, 2. 0 U Taq enzyme ; Thermal cycling was programmed by 1 cycle for 2 min at 94℃ , followed by 40 cycles for lmin at 94℃ 45see at 36℃, and 90 sec at 72 ℃ and finally, by 1 cycle for 10 rain at 72 ℃, storied at 4℃. Conclusion The optimized reaction system and program of RAPD for ginseng has established. |
| Keywords: | RAPD |
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