A simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification

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 10 8 Escherichia 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.

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Funding

This work was financially supported by the National Natural Science Foundation of China (81801264), National Key Research and Development Programs of China (2018YFE0113300) and the Key Project of Natural Science Foundation of Shandong Province (ZR2020KH030).

Author information

  1. Xiaoli Zhao and Yong Li contributed equally to this work.

Authors and Affiliations

  1. 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 Xiaoli Zhao, Yong Li, Yake Duan & Cuiping Ma
  2. Biology Department, College of Science, UAE University, Al Ain, 15551, UAE Amr Amin
  3. Biology Department, School of Sciences and Humanities, Nazarbayev University, Nur-Sultan, Kazakhstan, 010000 Yingqiu Xie
  4. 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 Chao Shi
  1. Xiaoli Zhao