Ivanov Artemii A, Tyapkin Alexandr V, Golubeva Tatiana S
Institute of Cytology and Genetics, Novosibirsk 630090, Russia.
Department of Natural Science, Novosibirsk State University, Novosibirsk 630090, Russia.
Curr Issues Mol Biol. 2023 Apr 18;45(4):3517-3524. doi: 10.3390/cimb45040230.
RNA isolation from fungi and fungus-like organisms is not an easy task. Active endogenous RNases quickly hydrolyze RNA after the sample collection, and the thick cell wall prevents inhibitors from penetrating the cells. Therefore, the initial collection and grinding steps may be crucial for the total RNA isolation from the mycelium. When isolating RNA from , we varied the grinding time of the Tissue Lyser and used TRIzol and beta-mercaptoethanol to inhibit the RNase. In addition, we tested the mortar and pestle grinding of mycelium in liquid nitrogen, with this method showing the most consistent results. During the sample grinding with the Tissue Lyser device, adding an RNase inhibitor proved to be a prerequisite, and the best results were achieved using TRIzol. We considered ten different combinations of grinding conditions and isolation methods. The classical combination of a mortar and pestle, followed by TRIzol, has proved to be the most efficient.
从真菌和类真菌生物中分离RNA并非易事。样本采集后，活跃的内源性核糖核酸酶会迅速水解RNA，而且厚厚的细胞壁会阻止抑制剂进入细胞。因此，最初的采集和研磨步骤对于从菌丝体中分离总RNA可能至关重要。在从[具体对象未提及]分离RNA时，我们改变了组织研磨仪的研磨时间，并使用TRIzol和β-巯基乙醇来抑制核糖核酸酶。此外，我们测试了在液氮中用研钵和研杵研磨菌丝体的方法，该方法显示出最一致的结果。在用组织研磨仪装置研磨样本的过程中，添加核糖核酸酶抑制剂被证明是一个先决条件，使用TRIzol可获得最佳结果。我们考虑了十种不同的研磨条件和分离方法组合。研钵和研杵的经典组合，随后使用TRIzol，已被证明是最有效的。
