软体动物分类学中的传统分子方法和组学技术越来越多地为软体动物的生物学提供信息。用于此类研究的 DNA 和 RNA 回收受到高度多样化软体动物的共同生物学特性的挑战。软体动物生物矿化、粘附结构和粘液涉及阻碍 DNA 提取或共纯化以抑制酶催化的分子过程的多酚蛋白和粘多糖。使用去污剂十六烷基三甲基溴化铵 (CTAB) 去除这些污染物的 DNA 提取方法重要地促进了软体动物的分子水平研究。软体动物色素可能会染色 DNA 样品并干扰分光光度法，因此需要凝胶电泳或荧光法进行准确定量。RNA 可以可靠地提取，但软体动物（如大多数原生动物）的 28S rRNA 中的“隐藏断裂”会导致 18S 和 28S rRNA 片段以电泳方式共同迁移。这对基于 18S 和 28S rRNA 比例的标准质量控制提出了挑战，该比例是为后口动物开发的。软体动物 rRNA 中的高 AT 含量阻止了多腺苷酸化 mRNA 的有效纯化。对这些问题的认识有助于分子疾病学的不断扩展，使博物馆标本和下一代测序的工作也成为可能，后者对 DNA 质量提出了前所未有的要求。从软体动物中提取核酸的替代方法可从文献中获得，重要的是，可从与研究软体动物分子生物学的其他人的交流中获得。

本文是 Theo Murphy 会议问题“软体动物基因组学：被忽视的门的广泛见解和未来方向”的一部分。

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Traditional molecular methods and omics-techniques across molluscan taxonomy increasingly inform biology of Mollusca. Recovery of DNA and RNA for such studies is challenged by common biological properties of the highly diverse molluscs. Molluscan biomineralization, adhesive structures and mucus involve polyphenolic proteins and mucopolysaccharides that hinder DNA extraction or copurify to inhibit enzyme-catalysed molecular procedures. DNA extraction methods that employ the detergent hexadecyltrimethylammoniumbromide (CTAB) to remove these contaminants importantly facilitate molecular-level study of molluscs. Molluscan pigments may stain DNA samples and interfere with spectrophotometry, necessitating gel electrophoresis or fluorometry for accurate quantification. RNA can reliably be extracted but the ‘hidden break’ in 28S rRNA of molluscs (like most protostomes) causes 18S and 28S rRNA fragments to co-migrate electrophoretically. This challenges the standard quality control based on the ratio of 18S and 28S rRNA, developed for deuterostome animals. High-AT content in molluscan rRNA prevents the effective purification of polyadenylated mRNA. Awareness of these matters aids the continuous expansion of molecular malacology, enabling work also with museum specimens and next-generation sequencing, with the latter imposing unprecedented demands on DNA quality. Alternative methods to extract nucleic acids from molluscs are available from literature and, importantly, from communications with others who study the molecular biology of molluscs.

This article is part of the Theo Murphy meeting issue ‘Molluscan genomics: broad insights and future directions for a neglected phylum’.