| 毛干DNA的快速提取及其对PCR的影响 |
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| 引用本文： | 管政,刘金钏,周宇,蒋小玲,杨曙明,陈爱亮. 毛干DNA的快速提取及其对PCR的影响[J]. 安徽农业科学, 2013, 0(25): 10232-10235 |
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| 作者姓名： | 管政 刘金钏 周宇 蒋小玲 杨曙明 陈爱亮 |
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| 作者单位： | 中国农业科学院农业质量标准与检测技术研究所,农业部农产品质量安全重点实验室,北京100081;中国农业科学院农业质量标准与检测技术研究所,农业部农产品质量安全重点实验室,北京100081;济宁医学院公共卫生学院,山东济宁272067 |
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| 基金项目： | 国家国际科技合作专项项目（2012DFA31140）;公益性行业（农业）科研专项经费（201203046）. |
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| 摘 要： | [目的]建立毛干DNA快速稳定的提取扩增策略,使这一简便取材能在畜牧、野生动物保护等采样困难的领域得以普及应用.[方法]采用PCR缓冲液快速提取及2轮PCR逐级稀释扩增的策略提取扩增黄牛毛干DNA微卫星位点.[结果]该方法可从低至0.1mg（约1 cm长）的毛干中提取得到DNA,并成功扩增核DNA微卫星位点.48份黄牛牛毛样本均成功提得DNA.72份扩增样本,除4份样本扩增效果较差,其余样品均能正常扩增.[结论]该研究方法可快速稳定的提取扩增目标基因,为毛干在相关领域的普及应用奠定了基础.
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| 关 键 词： | 毛干 DNA 提取 PCR |
A Fast Hair Shaft DNA Extraction Method and Its Influence on PCR Amplification |
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| Affiliation: | GUAN Zheng(Institute of Quality Standard ＆ Testing Technology for Agro-Products, CAAS, Beijing 100081) |
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| Abstract: | [Objective] To establish a fast and stable hair shaft DNA extraction and amplification strategy,and make it to use in animal husbandry and wild animals protection.[Method] An improved PCR buffer fast extraction method and 2 rounds of PCR strategy were adopted to extract and amplify DNA locus of cattle hair shaft.[Result] The result showed that this strategy even can extract nuDNA （nuclear DNA） and amplify its target gcnes from 0.1 mg （about 1 cm） hair shaft.It successfully extracted DNA from all 48 hair shaft samples of cattle.In amplification stage,68 of 72 amplification samples have got its target genes.[Conclusion] This method can extract nuDNA from hair shaft and amplify its target genes fast and stable,which lays a foundation for application of hair shaft in relevant field. |
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| Keywords: | Hair shaft DNA Extraction PCR |
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