Data from: Modified low-salt CTAB extraction of high-quality DNA from contaminant rich tissues

The increasing use of high-throughput sequencing platforms has made the isolation of pure, high molecular weight DNA a primary concern for studies of a diverse range of organisms. Purification of DNA remains a significant challenge in many tissue and sample types due to various organic and inorganic molecules that co-precipitate with nucleic acids. Molluscs, for example, contain high concentrations of polysaccharides which often co-precipitate with DNA and can inhibit downstream enzymatic reactions. We modified a low-salt CTAB (MoLSC) extraction protocol to accommodate contaminant rich animal tissues and compared this method to a standard CTAB extraction protocol and two commercially available animal tissue DNA extraction kits using oyster adductor muscle. Comparisons of purity and molecular integrity showed that our in-house protocol yielded genomic DNA generally free of contaminants and shearing, whereas the traditional CTAB method and some of the commercial kits yielded DNA unsuitable for some applications of massively parallel sequencing (MPS). Our open-source MoLSC protocol provides a cost-effective, scalable, alternative DNA extraction method that can be easily optimized and adapted for sequencing applications in other contaminant rich samples.

随着高通量测序平台（high-throughput sequencing platforms）的应用日益普及，获取纯净、高分子量的脱氧核糖核酸（DNA）已成为各类生物研究的核心关切。由于各类有机与无机分子会与核酸（nucleic acids）共沉淀，在多数组织及样本类型中，DNA纯化仍是一项显著的技术挑战。以软体动物（Molluscs）为例，其体内富含多糖（polysaccharides），这类物质常与DNA共沉淀，进而抑制下游酶促反应。我们对低盐十六烷基三甲基溴化铵（CTAB）提取方案进行改良，得到适配富含污染物动物组织的提取流程（命名为改良低盐CTAB（MoLSC）方案）；随后以牡蛎闭壳肌为实验样本，将该方案与标准CTAB提取方案以及两款市售动物组织DNA提取试剂盒展开对比。通过对DNA纯度与分子完整性的对比评估，我们发现自研方案所得的基因组DNA通常无污染物残留且未发生剪切断裂；而传统CTAB法与部分市售试剂盒提取的DNA，则无法满足部分大规模平行测序（massively parallel sequencing，MPS）应用的需求。我们开源的MoLSC方案提供了一种经济高效、可扩展的替代性DNA提取方法，可轻松针对其他富含污染物的样本进行优化适配，满足各类测序应用场景的需求。