| 刘皓1,2，刘青1，吴旭干1，何杰1，董鹏生1，李彤1，成永旭1,2.不同提取方法和不同组织对中华绒螯蟹
DNA提取效果的比较研究[J].广东农业科学,2014,41(24):147-150 |
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不同提取方法和不同组织对中华绒螯蟹
DNA提取效果的比较研究 |
| Comparative study on extraction of genome DNA fromdifferent tissues of Chinese mitten crab (Eriocheirsinensis) by dissimilar methods |
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| DOI： |
| 中文关键词: 中华绒螯蟹 DNA 提取 酚-氯仿法 试剂盒法 不同组织 |
| 英文关键词: Chinese mitten crabs (Eriocheir sinensis) DNA extraction phenol-chloroform method DNA extraction
kit different tissues |
| 基金项目:国家“863”计划项目（012AA10A409-5）；科技部
科技型中小企业技术创新项目（14C26213201214）；上海市科学
技术委员会项目（13231203504, 13DZ2280500）；上海高校水产
学一流学科建设项目（沪教科2012-62） |
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| 摘要点击次数: 1777 |
| 全文下载次数: 626 |
| 中文摘要: |
| 为了优化中华绒螯蟹基因组DNA 提取方法，采用酚-氯仿法和试剂盒法分别对中华绒螯蟹的鳃、肌肉、
血淋巴液和刚毛4 种组织进行DNA 提取，并从纯度、浓度、片段完整性及微卫星标记PCR 扩增效果4 个方面评价提
取DNA 的质量。结果显示院酚-氯仿法提取的DNA 片段比试剂盒法提取的更完整，且DNA 浓度较高，但纯度稍低曰
无论何种提取方法，刚毛和血淋巴液中的DNA 纯度都高于鳃和肌肉，4 种组织中的DNA 含量依次为鳃>肌肉>刚毛>
血淋巴液，未剪碎刚毛中未能提取到基因组DNA曰微卫星标记的电泳结果表明，两种提取方法下4 种组织中提取的
DNA 均可满足微卫星标记的应用要求。综上，本研究建立了一种简便的非损伤性中华绒螯蟹的采样及其DNA 提取
方法，这对于进行中华绒螯蟹遗传育种和分子标记研究等方面具有一定的积极作用。 |
| 英文摘要: |
| To optimize genome DNA extraction method of Chinese mitten crabs, genome DNA was extracted from gills,
muscle, blood and bristles of Chinese mitten crabs using phenol -chloroform method and DNA extraction kit, and the
quality of these DNA samples was evaluated in four aspects: the purity, concentration, fragment completeness and
microsatellite marker PCR amplification effect. The results showed that, compared with the method of DNA extraction kit,
the DNA extracted by phenol-chloroform method was much more complete, had higher concentration, but lower purity.
Both the extraction methods showed that the purity of DNA in bristles and blood were higher than that in the gills and
muscle, and the descending order of DNA content was gill, muscle, bristles, blood, while the uncutting bristles were failed
to extract genomic DNA. Microsatellite PCR electrophoresis results showed that the DNA extracted from four different
tissues by two dissimilar DNA extraction methods could meet the requirements of the application of microsatellite markers.
In conclusion, the present study established a convenient noninvasive sampling method and a procedure of DNA extraction
that could promote genetic breeding and molecular marker research of E. sinensis. |
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