Genomics Research Centre, Griffith Health Institute, Griffith University, Gold Coast Campus, Parklands Drive, Southport, QLD, 4215, Australia.
Mol Biol Rep. 2012 May;39(5):5961-6. doi: 10.1007/s11033-011-1408-8. Epub 2012 Jan 7.
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.
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。这些特征尚未针对某些实验室技术进行全面评估，例如 DNA 提取的盐析法，该方法已被排除在迄今为止发表的不同研究中的比较之外。我们比较了三种不同的方案（传统的盐析法、改良的盐析法和市售试剂盒法），以确定提取 DNA 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本中提取基因组 DNA，并比较了所获得产物的数量（提取的 DNA 浓度和每毫升使用血液获得的 DNA）和质量（260/280 比值和聚合酶链反应产物扩增）的结果。平均而言，所有三种方法在最终结果之间均无统计学显着差异，但当我们考虑每种方法的时间和成本时，它们显示出非常显着的差异。与商业试剂盒和传统盐析法相比，改良盐析法的成本分别降低了七倍和两倍，与传统盐析法相比，时间从 3 天缩短到 1 小时。这突出了改良盐析法作为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本时。
