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湖泊科学  2014 

两种提取枝角类休眠卵基因组DNA方法的比较

DOI: 10.18307/2014.0419

Keywords: 休眠卵,DNA提取,盔型溞,氯仿异戊醇提取法,玻璃乳提取法

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Abstract:

休眠卵是枝角类在环境胁迫条件下通过有性生殖形成的,休眠卵的孵化对种群补充与恢复有着重要作用.沉积于底泥中的休眠卵保存了不同阶段的种群遗传信息,有效地提取枝角类休眠卵基因组DNA是进一步研究水体枝角类遗传多样性的关键环节.采用目前国际上使用的氯仿异戊醇提取法和玻璃乳提取法分别对流溪河水库底泥中表层和底层的盔型溞(Daphniagaleata)休眠卵进行基因组DNA提取的比较分析.结果显示,氯仿异戊醇提取法成功率为27.5%,平均浓度为14.25±1.84ng/μl;玻璃乳提取法成功率为65.0%,平均浓度为28.37±2.56ng/μl.无论是提取成功率还是提取浓度玻璃乳提取法都显著高于氯仿异戊醇提取法,且玻璃乳提取法所用试剂少,不涉及有毒试剂,操作步骤简单,整个提取过程所用时间短,玻璃乳提取法提取枝角类休眠卵基因组DNA是一种快捷实用的方法.

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