A simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification
Zhao X. Li Y. Duan Y. Amin A. Xie Y. Shi C. Ma C.
November 2021Springer Science and Business Media Deutschland GmbH
Analytical and Bioanalytical Chemistry
2021#413Issue 266469 - 6477 pp.
RNA isolation from bacteria is technically difficult due to the RNA characteristic of labile and vulnerable degradation. Many reagents were explored for cellular lysis and complete inhibition of RNase. However, the available methods for RNA isolation are either of low efficiency or time-consuming. Here, we developed a rapid and accessible protocol for RNA isolation that combined a simplified cell lysis and RNA release by formamide-based solution and RNA purification by chitosan-modified silica membrane for the first time. With this method, we obtained about ~ 28 μg of total RNA from 108Escherichia coli cells. The entire procedure can be done within 15 min without redundant pipetting steps. The purity of extracted RNA was comparable to that of commercial kits, but the cost was much lower. Furthermore, the yielded RNA was successfully used in downstream enzymatic reactions, such as reverse transcription and quantitative real-time PCR. This new method would be of benefit for an extensive range of gene expression analyses in bacterial organisms. Graphical abstract: [Figure not available: see fulltext.]
Bacteria , Chitosan-modified silica membrane , Formamide , Nucleic acid extraction , RNA purification
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Shandong Provincial Key Laboratory of Biochemical Engineering, Qingdao Nucleic Acid Rapid Detection Engineering Research Center, College of Chemistry and Molecular Engineering, College of Marine Science and Biological Engineering, Qingdao University of Science and Technology, Qingdao, 266042, Shandong, China
Biology Department, College of Science, UAE University, Al Ain, 15551, United Arab Emirates
Biology Department, School of Sciences and Humanities, Nazarbayev University, Nur-Sultan, 010000, Kazakhstan
Qingdao Nucleic Acid Rapid Testing International Science and Technology Cooperation Base, College of Life Sciences, Department of Pathogenic Biology, School of Basic Medicine, and Department of Clinical Laboratory, The Affiliated Hospital of Qingdao University, Qingdao University, Qingdao, 266071, Shandong, China
Shandong Provincial Key Laboratory of Biochemical Engineering
Biology Department
Biology Department
Qingdao Nucleic Acid Rapid Testing International Science and Technology Cooperation Base
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