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Comparison of genomic DNA extraction techniques from whole blood samples: a time, cost and quality evaluation study

机译:从全血样本中提取基因组DNA的技术比较:时间,成本和质量评估研究

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Genomic DNA obtained from patient whole blood samples is a key element for genomic research. Advantages and disadvantages, in terms of time-efficiency, cost-effectiveness and laboratory requirements, of procedures available to isolate nucleic acids need to be considered before choosing any particular method. These characteristics have not been fully evaluated for some laboratory techniques, such as the salting out method for DNA extraction, which has been excluded from comparison in different studies published to date. We compared three different protocols (a traditional salting out method, a modified salting out method and a commercially available kit method) to determine the most cost-effective and time-efficient method to extract DNA. We extracted genomic DNA from whole blood samples obtained from breast cancer patient volunteers and compared the results of the product obtained in terms of quantity (concentration of DNA extracted and DNA obtained per ml of blood used) and quality (260/280 ratio and polymerase chain reaction product amplification) of the obtained yield. On average, all three methods showed no statistically significant differences between the final result, but when we accounted for time and cost derived for each method, they showed very significant differences. The modified salting out method resulted in a seven- and twofold reduction in cost compared to the commercial kit and traditional salting out method, respectively and reduced time from 3 days to 1 hour compared to the traditional salting out method. This highlights a modified salting out method as a suitable choice to be used in laboratories and research centres, particularly when dealing with a large number of samples.
机译:从患者全血样本中获得的基因组DNA是基因组研究的关键要素。在选择任何特定方法之前,需要考虑可用于分离核酸的程序在时间效率,成本效益和实验室要求方面的优缺点。这些特性尚未在某些实验室技术中得到充分评估,例如用于DNA提取的盐析法,迄今为止,在已发表的不同研究中均未将其排除在比较范围之外。我们比较了三种不同的方案(传统的盐析方法,改良的盐析方法和市售试剂盒方法),以确定提取DNA的最具成本效益和时间效率的方法。我们从乳腺癌患者志愿者的全血样品中提取了基因组DNA,并比较了所获得产品的数量(提取的DNA浓度和每毫升所用血液的DNA浓度)和质量(260/280的比例和聚合酶链)的结果反应产物的扩增)。平均而言,所有三种方法在最终结果之间均没有统计学上的显着差异,但是当我们考虑每种方法的时间和成本时,它们表现出非常显着的差异。改进的盐析方法与商用套件和传统的盐析方法相比,成本分别降低了七倍和两倍,并且与传统的盐析方法相比,时间从3天减少至1小时。这突出了一种改进的盐析方法,是在实验室和研究中心使用的合适选择,特别是在处理大量样品时。

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