凡纳滨对虾(Litopenaeus vannamei)组织样品常温保存液的筛选和优化

Selection and Optimization of Simple and Convenient Sample Solutions for Shrimp Tissue Preservation at Normal Temperature

中文关键词: 铵盐  乙醇  常温  RNA

英文关键词: Ammonium salt  Ethanol  Normal temperature  RNA

中文摘要:

      以不同浓度乙醇为溶剂，配制不同pH的硫酸铵和乙酸铵饱和溶液，测试其对凡纳滨对虾组织的渗透效应和在常温条件下对RNA的保护作用。因为乙酸铵在乙醇中的溶解度远高于硫酸铵，且其铵离子的摩尔浓度在0−80%的乙醇溶剂中比较稳定。研究结果显示，乙酸铵比硫酸铵能更快渗透到整体的对虾组织中，并达到更高的浓度。50%、70%及以上的乙醇为溶剂的饱和硫酸铵、50%乙醇为溶剂的饱和乙酸铵(pH=6.0)溶液保存RNA的完整性较好，RNA拷贝数较大，半衰期较长。综合分析，推荐50%乙醇的饱和乙酸铵溶液(pH=6.0)作为对虾组织样品的常温保存液，该结果为野外样品采集及常温运输提供了一种有效方法。

英文摘要:

      Molecular biology technologies have been widely used in aquatic animal disease diagnosis and molecular pathology studies, which require high quality of nucleic acid. However, how to preserve sample quality at normal temperature is rarely investigated. In this study, we compared the preserving effect of RNA in the tissues of shrimp Litopenaeus vannamei stored in different preservation solutions at normal temperature to find one preservation solution that can be used at normal temperature and long-distance transportation. Saturated solutions of ammonium sulfate and ammonium acetate in a serial of solvents with different ethanol concentrations were used to preserve shrimp tissues. The results showed that ammonium acetate has higher solubility in ethanol solution than ammonium sulfate. The molarity of ammonium ion of ammonium acetate is steadier in 0−80% ethanol. Ammonium acetate penetrates into the tissue of intact shrimp faster and reaches a higher concentration than ammonium sulfate does. RNA with better integrity, higher copies, and longer half-life period was achieved in the tissue preserved in more than 70% ethanol, saturated ammonium sulfate in the solvent of 50% and 70% ethanol, saturated ammonium acetate in the solvent of 50% ethanol. These results suggest that the saturated ammonium acetate in 50% ethanol as the solvent (pH=6.0) can be used as sample preserving solution to preserve RNA in shrimp tissues at normal temperature.