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Tomoko Matsuda 2021. RNA isolation from suspended animal cells, cDNA library construction, and RNA-Seq for gene expression analysis. protocols.io https://dx.doi.org/10.17504/protocols.io.57ng9meCopy Citation Copied
URL: https://dx.doi.org/DOI:10.17504/protocols.io.57ng9me
Authors: Tomoko Matsuda
Summary: This protocol is for RNA Isolation from suspended Chinese hamster ovary (CHO) cells. Wash cells once in 1× PBS and isolate RNA using Qiashredder columns and the RNeasy mini kit. In the case of RNA-Seq, which reveals the presence and quantity of RNA in a biological sample at any moment, it is necessary to consider that gene expression responds over a short time interval (several seconds to a few minutes) in many organisms. Therefore, to isolate RNA that accurately reflects the transcriptome at the point of harvest, raw biological samples should be processed by lysing and homogenizing in RNA lysis buffer containing guanidine thiocyanate (buffer RLT, RNA lysis buffer, QIAGEN) as soon as possible (within 15 minutes, if possible).
Affiliations: Nihon BioData Corporation
Version: 1
Publication Date: 2021
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