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Updated: Jun 17, 2025
RNA Isolation of Pseudomonas aeruginosa Colonizing the Murine Gastrointestinal Tract
Published on: September 28, 2011
Effective RNA isolation method for gram-positive and acid-fast bacteria: metamorphosed from conventional RNA
Jignasa H Bera1, Leyon Selvin Raj A1, Hemant Kumar2
1Department of Medical Laboratory Technology, Bapubhai Desaibhai Patel Institue of Paramedical Sciences (BDIPS), Charotar University of Science and Technology, CHARUSAT campus, Changa, Anand, Gujarat, 388421, India.
Abstract:
The RNA-based study provides an excellent indication of an organism's gene expression profile. Obtaining high-yield and high-purity RNA from Gram-positive and acid-fast bacteria is difficult without high-end kits and facilities. We optimised effective and simple protocol for RNA isolation that is a combination of enzymatic, physical and chemical treatment to disrupt cells. We successfully isolated high quality intact total RNA with yields ranging from 23.13 ± 0.40 to 61.51 ± 0.27 µg and the 260/280 purity ratio of 1.95 ± 0.01 to 2.05 ± 0.01 from Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, and Mycobacterium smegmatis. These results represents a significantly enhanced yield and purity compared to other combination of techniques which we performed. Compared to previous studies the yield obtained by this method is high for the studied organisms. Furthermore the yielded RNA was successfully used for downstream applications such as quantitative real time PCR. The described method can be easily optimised and used for various bacteria.
