| 引用本文： |
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金忠民,沙伟,孙天国,孙雪巍.蓝萼香茶菜随机扩增多态DNA(RAPD)反应体系优化[J].广西科学,2004,11(3):258-260. [点击复制]
- Jin Zhongmin,Sha Wei,Sun Tianguo,Sun Xuewei.Optimization of RAPD System of Isodon japonica (Burm.f.) Hara var.glaucocalyx (Maxim.) Hara[J].Guangxi Sciences,2004,11(3):258-260. [点击复制]
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| 摘要: |
| 试验以香茶菜属植物的蓝萼香茶菜[Isodon japonica(Burm.f.) Hara var.glaucocalyx(Maxim.)]为材料,采用改进的CTAB法提取基因组DNA,首次建立适合于蓝萼香茶菜的RAPD(Random amplified polymorphic NDA,随机扩增的多态性)分析的PCR反应体系。通过对RAPD条件优化分析,确定蓝萼香茶菜RAPD的最佳方案为:25μl反应体系中包括三磷酸脱氧核苷酸(dNTPs)0.1mmol/L,引物0.4μmol/L,模板DNA 90ng,Taq酶(Sangon)3u,2.5μl 10×buffer缓冲液(含20mmol/L Mg2+),其余部分用无菌超纯水补平;PCR扩增循环为93℃预变性4min,93℃变性45s,36℃复性75s,72℃延伸2min,40个循环,最后72℃延伸10min。 |
| 关键词: 蓝萼香茶菜 RAPD 优化 |
| DOI： |
| 投稿时间：2004-01-29 |
| 基金项目:齐齐哈尔市科技局计划项目。 |
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| Optimization of RAPD System of Isodon japonica (Burm.f.) Hara var.glaucocalyx (Maxim.) Hara |
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Jin Zhongmin, Sha Wei, Sun Tianguo, Sun Xuewei
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| (The Life Science and Engineering College, Qiqihaer University, Qiqihaer, Heilongjiang, 161006, China) |
| Abstract: |
| The total DNA from the young leaves of I.japonica (Burm.f.) Hara var.glaucocalyx (Maxim.) Hara was successfully extracted by using the developed CTAB method and a steady system of PCR reaction for I.japonica(Burm.f.) Hara var.glaucocalyx (Maxim.) Hara was built.The optimal PCR system for RAPD analysis was as follows:0.1 mmol/L dNTPs,0.4 μmol/L random primer,90ng DNA template,3u Taq polymerase in 25 μl reaction solution.The reaction program was devised for one cycle at 93℃ 4 minutes and followed by 40 cycles, each with 45 seconds at 93℃,75 seconds at 37℃,2 minutes at 72℃,and a final extension at 72℃ for 10 minutes. |
| Key words: Isodon japonica(Burm.f.) Hara var.glaucocalyx(Maxim.) Hara RAPD optimization |
