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。
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. Conclusions： 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.
出处
《现代生物医学进展》
CAS
2015年第6期1138-1140,共3页
Progress in Modern Biomedicine
关键词
基因组DNA
提取
外周血
Genomic DNA
Extraction
Peripheral blood
