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Abstract

Biochemistry

大鼠胰腺组织快速、经济高效的RNA提取

Published: September 19th, 2020

DOI:

10.3791/61255

1Department of Biochemistry, School of Medicine, Shiraz University of Medical Sciences, 2Endocrinology and Metabolism Research Center, Nemazee Hospital, Shiraz University of Medical Sciences, 3English Language Department, Faculty of Paramedical Sciences, Shiraz University of Medical Sciences, 4Department of e-Learning in Medical Sciences, Virtual School and Center of Excellence in e-Learning, Shiraz University of Medical Sciences, 5Autophagy Research Center, School of Medicine, Shiraz University of Medical Sciences

Abstract

无论提取方法如何,组织和细胞系的优化RNA提取分四个阶段进行:1) 均质化,2) RNA中蛋白质的有效变性,3) 核糖核糖核素灭活,以及 4) 去除DNA、蛋白质和碳水化合物中的污染。然而,当组织中有高水平的RNA时,保持RNA的完整性是非常费力的。自发的自解使得在不损害胰腺组织的情况下从胰腺组织中提取RNA变得非常困难。因此,在提取过程中需要一种实用的RNA提取方法来维持胰腺组织的完整性。通过在不到2分钟内获得20-30毫克大鼠胰腺组织并提取RNA,对现有方案进行了实验和比较研究。结果通过电泳评估。实验进行了三次,以概括结果。当RNA提取试剂用作试剂时,在-80°C下将胰腺组织浸入RNA稳定试剂中24小时,产生高完整性RNA。所得结果与具有自旋柱结合的商业套件所得结果相当。

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