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

Xiaoli Zhao, Yong Li, Yake Duan, Amr Amin, Yingqiu Xie, Chao Shi, Cuiping Ma

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

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.]

Original languageEnglish
Pages (from-to)6469-6477
Number of pages9
JournalAnalytical and Bioanalytical Chemistry
Volume413
Issue number26
DOIs
Publication statusPublished - Nov 2021

Keywords

  • Bacteria
  • Chitosan-modified silica membrane
  • Formamide
  • Nucleic acid extraction
  • RNA purification

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

Fingerprint

Dive into the research topics of 'A simple methodology for RNA isolation from bacteria by integration of formamide extraction and chitosan-modified silica purification'. Together they form a unique fingerprint.

Cite this