SHI Kai-ming,DENG Zhi-jun,FANG Xiang-liang.Study on Extraction of Genome DNA in Gongshui Pomelo[J].Northern Horticulture,2014,38(21):97-100.
贡水白柚基因组DNA提取方法研究
- Title:
- Study on Extraction of Genome DNA in Gongshui Pomelo
- 文章编号:
- 1001-0009（2014）21-0097-04
- Keywords:
- Gong-shui pomelo(Citrus grandis(L.) Osbeck); DNA; extraction
- 分类号:
- S 602
- 文献标志码:
- A
- 摘要:
- 以“贡水白柚”为试材，采用盐乙醚处理法、苯酚氯仿处理法、基本CTAB处理法3种DNA提取方法来处理“贡水白柚”基因组DNA，研究不同提取方法对DNA样品的产率、外观、酶切电泳等方面的影响。结果表明：用盐乙醚法所提DNA外观呈纯白色，冷冻沉淀为纤维状，效果最好；用苯酚氯仿法处理幼穗期柚叶片基因组提取DNA产率最高，DNA浓度达118 ng/μL；EcoRⅠ酶切图谱用CTAB处理法和盐乙醚处理法效果均较好，苯酚氯仿处理法则拖尾严重，效果差；RAPD引物PCR扩增效果图表明，3种方法均能扩增出清晰条带，均适合于有关“贡水白柚”基因组的相关研究。
- Abstract:
- Taking Gongshui pemolo as test materials,water-saturated ether method，DNA phenol/chloroform method and basic CTAB method were used to extract DNA genome of Gongshui pomelo,the analysis and comparison was conducted from the yield of DNA samples,the appearance of DNA samples and the enzyme electrophoresis of DNA samples.The results showed that the water-saturated ether method proposed the best effect of appearance,which was pure white and fibrous for the frozen precipitation.The yield of genome DNA was the highest from the tender spike with phenol/chloroform treatment of young pomelo leaves,which obtained DNA concentration of 118 ng/μL.It indicated that the quality of DNA was more better with basic CTAB method and water-saturated ether method than phenol/chloroform method by EcoRⅠ.moreover,three methods all amplified clear bands of PCR by RAPD primers.
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备注/Memo
第一作者简介：石开明（1980-），男，博士，讲师，现主要从事林学园艺植物遗传育种等研究工作。E-mail:skm331@163.com.
基金项目：湖北省教育厅资助项目（Q20122902）；国家自然科学基金青年科学基金资助项目（81303169）。
收稿日期：2014-05-19