| 不同保存条件下云杉八齿小蠹总DNA提取方法的比较分析 |
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| 引用本文： | 张晟铭,张真,王鸿斌,迟德富.不同保存条件下云杉八齿小蠹总DNA提取方法的比较分析[J].安徽农业科学,2009,37(13):5883-5885. |
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| 作者姓名： | 张晟铭 张真 王鸿斌 迟德富 |
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| 作者单位： | 东北林业大学生命科学学院,黑龙江哈尔滨,150040;中国林业科学研究院森林生态环境与保护研究所国家林业局森林保护学重点实验室,北京,100091;中国林业科学研究院森林生态环境与保护研究所国家林业局森林保护学重点实验室,北京,100091;东北林业大学生命科学学院,黑龙江哈尔滨,150040 |
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| 基金项目： | 国际科技合作基金项目(2006DFA31790); 黑龙江省重大科技攻关项目(GB06B304) |
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| 摘 要： | 目的]探讨云杉八齿小蠹总DNA的提取方法。方法]将野外采集的云杉八齿小蠹分别保存在95％的酒精浸泡、自然风干和-40℃条件下，采用改良CTAB法、KAc法、盐析法和SDS法对不同保存条件下样品材料总DNA进行提取。结果]结果表明，4种方法均可从95％酒精浸泡标本和-40℃冻存样品中提取出高质量的总DNA，相比之下改良CTAB法最为简单、高效、经济。以COI的通用引物对所提取的总DNA进行扩增检测，结果表明，采用改良的CTAB法可以从95％的酒精浸泡样品和-40℃冰存样品DNA中扩增出清晰的单一条带，片段大小700bp，经与已知昆虫的COI片段分子量对比，可以初步断定其为COI片段。结论]该研究结果为今后小蠹类的分子系统学研究奠定基础。
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| 关 键 词： | 云杉八齿小蠹 DNA提取 COI序列 |
Comparative Analysis on Methods for Genomic DNA Extraction of Ips typographus L. |
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| ZHANG Sheng-ming et al.Comparative Analysis on Methods for Genomic DNA Extraction of Ips typographus L.[J].Journal of Anhui Agricultural Sciences,2009,37(13):5883-5885. |
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| Authors: | ZHANG Sheng-ming |
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| Institution: | ZHANG Sheng-ming et al (Life Science College of Northeast Forestry University,Haerbin,Heilongjiang 150040) |
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| Abstract: | Objective]The purpose of this research was to discuss methods for genomic DNA extraction of Ips typographus L..Method] Using four methods,inculding improved cetyltrimethyl ammonium bromide(CTAB) method,the potassium acetate(KAc) method,the salting-out method and the sodium dodecyl sulfate(SDS) method extracted the genomic DNA of Ips typographus L.preserved in 95% ethanol,naturally dried or frozen under-40 ℃ conditions.Result]The results indicated that all of the four methods could extract the purity and quantity genomic DNA from beetles dipped in 95% ethanol preserved and frozen ones,and the improved CTAB method was the most easy,efficient and ecomomic.Using the obtained DNA and the universal COI primer could amplify a clear single band,about 700 bp sequence.Compared the molecular weight with another insects' COI sequence,a deduction could be draw that it was the COI sequence of I.typographus L..Conclusion]This study will provide a foundation research for the molecular systematics of I.typographus L.. |
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| Keywords: | Ips typographus L DNA extraction COI Sequence |
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