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Effective RNA isolation method for gram-positive and acid-fast bacteria: metamorphosed from conventional RNA isolation techniques.
- Source :
-
Archives of microbiology [Arch Microbiol] 2024 Aug 07; Vol. 206 (9), pp. 369. Date of Electronic Publication: 2024 Aug 07. - Publication Year :
- 2024
-
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.<br /> (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)
- Subjects :
- Staphylococcus aureus genetics
Staphylococcus aureus isolation & purification
Gram-Positive Bacteria genetics
Gram-Positive Bacteria isolation & purification
Staphylococcus epidermidis genetics
Staphylococcus epidermidis isolation & purification
Enterococcus faecalis genetics
Enterococcus faecalis isolation & purification
Mycobacterium smegmatis genetics
RNA, Bacterial genetics
RNA, Bacterial isolation & purification
Subjects
Details
- Language :
- English
- ISSN :
- 1432-072X
- Volume :
- 206
- Issue :
- 9
- Database :
- MEDLINE
- Journal :
- Archives of microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 39110213
- Full Text :
- https://doi.org/10.1007/s00203-024-04077-2