| 王 丹 邓春青△ 赵龙凤.两种外周血 DNA 提取方法的比较[J].,2015,15(6):1138-1140 |
| 两种外周血 DNA 提取方法的比较 |
| Comparison of Two Kinds of Peripheral Blood DNA Extraction Methods |
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| DOI： |
| 中文关键词: 基因组 DNA 提取 外周血 |
| 英文关键词: Genomic DNA Extraction Peripheral blood |
| 基金项目: |
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| 中文摘要: |
| 目的： 外周血 DNA 的提取是研究乙型肝炎病毒相关临床疾病的基础， 所提取 DNA 的质与 量直接关乎下游研究的成败， 经
济、高效、便捷的外周血 DNA 提取方法对于疾病分子水平的研究尤为 重要， 本实验旨 在比较两种外周血 DNA 提取方法， 从而为
临床研究提供有力的参考。 方法： 以外周抗凝血为试验样本，分别采用 改良盐析法和 DNA 提取试剂盒法（硅胶柱纯化）进行基因
组 DNA 的提取，通过分光光度仪测量 DNA 浓度和纯度， 并进行 PCR 扩增及电泳实验。 比较改良盐析法与 试剂盒提取法（硅胶柱
纯化）的效果。 结果： 试剂盒提取法（硅胶柱纯化）标本用 量甚微，省时，提取 DNA 纯度高，步骤繁琐， PCR 条带单一、亮度差；改良
盐析法操作步骤少，提取 DNA 浓度高， PCR 条带亮度佳、杂带多 ， 耗时长。 结论： 两组方法各有优缺点， 试剂盒提取法（硅胶柱纯
化）可靠、快速， 但所获 DNA 量少、极易降解， 改良盐析法耗时， 但所获 DNA 浓度高、量多 ， 可根据实验时间 与 经费， 实验所需的
DNA 纯度与 浓度， 提供的样本体积等不同 的临床研究需求及条件来综合选择适宜的提取方法。 |
| 英文摘要: |
| Objective:Extracted DNA from peripheral blood of hepatitis B virus-related clinical disease is the basis of researching,
the quality and quantity of DNA extracted directly related to the success of downstream research. Economic, efficient and convenient
peripheral blood DNA extraction method for the study of disease at the molecular level is particularly important. Our study was designed
to compare the two kinds of peripheral blood DNA extraction method, thereby providing a strong reference for clinical research.Methods:Anticoagulation was used for the test sample. Improved salting-out method and DNA extraction kit method (purification by
silica gel column) were used for genomic DNA extraction respectively, by spectrophotometer meter measured the DNA concentration
and purity, PCR amplification and electrophoresis experiment was carried on. Comparison of the effect of improved salting-out and DNA
extraction kit.Results:Kit (purification by silica gel column), which saved time, with higher DNA-purity and better DNA single PCR
bands,had little dosage of extraction specimens. Improved salting-out method had easier operation and higher DNA-yield, but the PCR
band included many impurity bands.Conclusion:Both the two methods had advantages and disadvantages, kit (purification by silica gel
column) was reliable and fast, but received less DNA which was degraded easily, improved salting was time-consuming, but received
higher DNA concentrations and the amount. According to different clinical research needs and conditions, for example, the experimental
time and money, DNA purity and concentration required for the experiment, different volumes of the sample to provide comprehensive
selection of suitable extraction method. |
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