| 流式细胞术检测4种提取沙棘核DNA方法的比较 |
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| 引用本文： | 周香艳,张宁,王旺田,王翠玲,吴兵.流式细胞术检测4种提取沙棘核DNA方法的比较[J].甘肃农业大学学报,2012,47(4):155-160. |
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| 作者姓名： | 周香艳 张宁 王旺田 王翠玲 吴兵 |
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| 作者单位： | 甘肃农业大学生命科学技术学院,甘肃兰州,730070 |
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| 摘 要： | 采用4种不同提取细胞核的方法,测定了6个不同沙棘品种的核DNA含量,比较了4种不同提取方法测得的CV值的差异.结果表明:4种提取沙棘核DNA的方法所测得的内标CV值为2.49%～11.31%,待测样CV值为2.53%～12.08%.当Tris.MgCl2法测定时,细胞核全部破碎,故未得到流式图;用Otto's法测得的CV值最小,内标与待测样CV值分别为2.49%～3.35%,2.53%～3.56%.因此,Otto's法为最适宜提取沙棘细胞核DNA的方法.
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| 关 键 词： | 沙棘 流式细胞术 细胞核提取液 核DNA含量 |
Comparison of different nuclear isolation buffers for species of Hippophae DNA flow cytometry |
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| ZHOU Xiang-yan
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ZHANG Ning
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WANG Wang-tian
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WANG Cui-ling
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WU Bing.Comparison of different nuclear isolation buffers for species of Hippophae DNA flow cytometry[J].Journal of Gansu Agricultural University,2012,47(4):155-160. |
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| Authors: | ZHOU Xiang-yan ZHANG Ning WANG Wang-tian WANG Cui-ling WU Bing |
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| Institution: | (College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730000,China) |
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| Abstract: | The nuclear DNA content of six different species of Hippophae were detected using four nuclear isolation buffers(Galbraith’s,LB01,Otto’s and Tris·MgCl2),comparing the difference of CV in the four buffers,so as to find the optimum method for lysing nuclear DNA in species of Hippophae,the standard CV were measured ranging from 2.49% to 11.31% and the samples CV varied from 2.53% to 12.08%.Histograms of relative fluorescence intensity were not obtained using Tris·MgCl2 for a large number of debris emerging,but Otto’s was the optimum buffer(the standard CV=2.49%~3.35%%;the samples CV= 2.53%~3.56%) for preparing suspension of intact nuclei of Hippophae.The results might provide guidance for the future experiments of other plants with similar ploidy and cell component,and promote applications of flow cytometry in plant research. |
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| Keywords: | Hippophae flow cytometry nuclear isolation buffer nuclear DNA content |
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