College of Horticulture, Gansu Agricultural University, Lanzhou, 730070, People's Republic of China.
Sci Rep. 2022 Feb 18;12(1):2825. doi: 10.1038/s41598-022-06810-7.
The high quality, yield and purity total RNA samples are essential for molecular experiments. However, harvesting high quality RNA in Lilium davidii var. unicolor is a great challenge due to its polysaccharides, polyphenols and other secondary metabolites. In this study, different RNA extraction methods, namely TRIzol method, the modified TRIzol method, Kit method and cetyltrimethylammonium bromide (CTAB) method were employed to obtain total RNA from different tissues in L. davidii var. unicolor. A Nano drop spectrophotometer and 1% agarose gel electrophoresis were used to detect the RNA quality and integrity. Compared with TRIzol, Kit and CTAB methods, the modified TRIzol method obtained higher RNA concentrations from different tissues and the A260/A280 ratios of RNA samples were ranged from 1.97 to 2.27. Thus, the modified TRIzol method was shown to be the most effective RNA extraction protocol in acquiring RNA with high concentrations. Furthermore, the RNA samples isolated by the modified TRIzol and Kit methods were intact, whereas different degrees of degradation happened within RNA samples isolated by the TRIzol and CTAB methods. In addition, the modified TRIzol method could also isolate high-quality RNA from other edible lily bulbs. Taken together, the modified TRIzol method is an efficient method for total RNA isolation from L. davidii var. unicolor.
高质量、高产量和高纯度的 RNA 样品是分子实验的关键。然而,由于百合属杂种紫斑百合中含有多糖、多酚和其他次生代谢物,因此从该植物中提取高质量的 RNA 是一项巨大的挑战。在本研究中,我们采用了不同的 RNA 提取方法,即 TRIzol 法、改良的 TRIzol 法、试剂盒法和溴化十六烷基三甲铵(CTAB)法,从百合属杂种紫斑百合的不同组织中提取总 RNA。使用 Nano 微量分光光度计和 1%琼脂糖凝胶电泳检测 RNA 的质量和完整性。与 TRIzol 法、试剂盒法和 CTAB 法相比,改良的 TRIzol 法从不同组织中获得了更高浓度的 RNA,RNA 样品的 A260/A280 比值范围为 1.97 至 2.27。因此,改良的 TRIzol 法是获取高浓度 RNA 的最有效 RNA 提取方案。此外,改良的 TRIzol 法和试剂盒法分离得到的 RNA 样品完整,而 TRIzol 法和 CTAB 法分离得到的 RNA 样品则有不同程度的降解。此外,改良的 TRIzol 法还可以从其他可食用的百合鳞茎中分离出高质量的 RNA。综上所述,改良的 TRIzol 法是从百合属杂种紫斑百合中提取总 RNA 的有效方法。