| 西伯利亚蝗基因组DNA提取及RAPD分析条件的优化 |
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| 引用本文： | 葛风伟,季荣,曾献春,王婷,王小平.西伯利亚蝗基因组DNA提取及RAPD分析条件的优化[J].昆虫知识,2007,44(4):505-508. |
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| 作者姓名： | 葛风伟 季荣 曾献春 王婷 王小平 |
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| 作者单位： | 1. 新疆师范大学生命与化学学院,乌鲁木齐,830054
2. 新疆农业职业技术学院,昌吉州831100 |
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| 基金项目： | 国家自然科学基金项目(30460028)，新疆维吾尔自治区高校科研资助项目(XJEDU2005I23,XJEDU2004S20) |
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| 摘 要： | 以西伯利亚蝗Gomphocerus sibiricus(L.)为研究材料,利用改良的SDS法提取高质量的DNA,分别测试了dNTP浓度、镁离子浓度、TaqDNA聚合酶用量、模板DNA的量等因素对反应结果的影响。通过各因子的组合比较,建立了西伯利亚蝗RAPD优化体系:25μLPCR反应体系,10×buffer2·5μL;dNTP0·24mmol/L;MgCl22·0mmol/L;Taq DNA聚合酶1U;DNA模板45ng;引物30ng。扩增程序为:94℃预变性1min45s、94℃变性30s、35℃退火1min30s、72℃延伸2min,45个循环、72℃延伸10min。结果表明,利用优化的反应条件进行西伯利亚蝗基因组DNA分析,实验有着良好的重复性和稳定性。
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| 关 键 词： | 随机扩增多态性DNA 基因组DNA 西伯利亚蝗 巴里坤地区 |
| 修稿时间： | 2006-09-03 |
Extraction of genomic DNA and optimization of RAPD system for Gomphocerus sibiricus |
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| GE Feng-Wei,JI Rong,ZENG Xian-Chun,WANG Ting,WANG Xiao-Ping.Extraction of genomic DNA and optimization of RAPD system for Gomphocerus sibiricus[J].Entomological Knowledge,2007,44(4):505-508. |
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| Authors: | GE Feng-Wei JI Rong ZENG Xian-Chun WANG Ting WANG Xiao-Ping |
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| Institution: | 1. College of Life and Chemisty, Xinjiang Normal University, Urumqi 830054,China;2. Institute of Agricultural Professional Technology,Changji 831100,China |
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| Abstract: | By using an improved SDS method,we obtained high qualified genomic DNA of Gomphocerus sibiricus(L.).The effects of several parameters(contents of dNTP,Mg2+,Taq DNA polymerase and template DNA) on amplification were examined.The optimal reaction system of RAPD of G.sibiricus was determined as followed:0.24 mmol/L dNTP,2.0 mmol/L Mg2+,1.0 U Taq DNA polymerase,45 ng template DNA and 30 ng primer in total 25 μL reaction volume.Modified thermal profile consisted of an initial denaturation step at 94℃ for 1 min and 45 s,followed by 45 cycles of 94℃ for 30 s,35℃ 1 min and 30 s and 72℃ for 2 min,and a final exposure to 72℃ for 10 min. |
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| Keywords: | RAPD(random amplified polymorphic DNA) genomic DNA Gomphocerus sibiricus Balikun area |
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