Department of Infectious Disease, St Jude Children's Research Hospital, Memphis, Tennessee, USA.
Department of Pathology, St Jude Children's Research Hospital, Memphis, Tennessee, USA.
Biotechniques. 2019 Jun;66(6):285-289. doi: 10.2144/btn-2019-0016. Epub 2019 May 24.
A major challenge for microbiome studies is maintaining an even and accurate DNA extraction in the presence of samples with a wide range of bacterial content. Here we compare five DNA extraction methods using replicate stool samples that were diluted to create high and low biomass samples. Our results indicate greater variation in microbiome composition between high and low biomass samples than variation between methods. Many of the extraction methods had reduced yield from low biomass samples; however, our adapted plate column-based extraction method was evenly efficient and captured the largest number of high-quality reads. Based on these results, we have identified a DNA extraction method that ensures adequate yield in metagenomic microbiome studies that have samples with a broad range of bacterial content.
微生物组研究的一个主要挑战是在存在细菌含量范围广泛的样本的情况下保持均匀和准确的 DNA 提取。在这里,我们使用复制的粪便样本比较了五种 DNA 提取方法,这些样本被稀释以创建高和低生物量样本。我们的结果表明,高和低生物量样本之间的微生物组组成变化大于方法之间的变化。许多提取方法从低生物量样本中减少了产量;然而,我们改良的基于平板柱的提取方法均匀有效,捕获了最多数量的高质量读数。基于这些结果,我们已经确定了一种 DNA 提取方法,该方法可确保在具有广泛细菌含量的宏基因组微生物组研究中获得足够的产量。