State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangdong Provincial Key Laboratory of Ophthalmology & Visual Science, Guangzhou, 510060, China.
Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou, 510060, China.
Biotechniques. 2023 May;74(5):203-209. doi: 10.2144/btn-2022-0120. Epub 2023 May 26.
High-quality RNA isolation from recalcitrant adipose tissue with high lipid content and low cell numbers is difficult. Many studies have made efforts to optimize methods for isolating RNA from adipose tissue through combinations of column-based kits and phenol-chloroform methods, or through in-house protocols. However, the considerable complexity of these protocols and the various kits/materials required hamper their wide use. Herein, we describe an optimized protocol based on TRIzol reagent, which is the most accessible ready-to-use reagent for nucleic acid and/or protein isolation in laboratories. This article provides a step-by-step protocol yielding sufficient and qualified RNA from lipid-rich specimens for downstream applications.
从富含脂质和细胞数量少的顽固脂肪组织中分离高质量的 RNA 非常困难。许多研究通过柱试剂盒和苯酚-氯仿方法的组合，或者通过内部协议，都在努力优化从脂肪组织中分离 RNA 的方法。然而，这些方案的相当复杂性以及所需的各种试剂盒/材料阻碍了它们的广泛使用。在此，我们描述了一种基于 TRIzol 试剂的优化方案，TRIzol 试剂是实验室中最容易获得的用于核酸和/或蛋白质分离的即用型试剂。本文提供了一个分步方案，可从富含脂质的标本中获得足够和合格的 RNA，用于下游应用。
