Effective RNA isolation method for gram-positive and acid-fast bacteria: metamorphosed from conventional RNA isolation techniques.

RNA, Bacterial / genetics Staphylococcus aureus / genetics Gram-Positive Bacteria / genetics Staphylococcus epidermidis / genetics Enterococcus faecalis / genetics Mycobacterium smegmatis / genetics
Acid-fast bacteria Gram-positive bacteria Hard-to-lyse Quantitative real-time PCR RNA isolation

Journal

Archives of microbiology
ISSN: 1432-072X
Titre abrégé: Arch Microbiol
Pays: Germany
ID NLM: 0410427

Informations de publication

Date de publication:
07 Aug 2024
Historique:
received: 15 02 2024
accepted: 02 07 2024
medline: 7 8 2024
pubmed: 7 8 2024
entrez: 7 8 2024
Statut: epublish

Résumé

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.

Identifiants

DOI: 10.1007/s00203-024-04077-2 PMID: 39110213
pubmed: 39110213
doi: 10.1007/s00203-024-04077-2
pii: 10.1007/s00203-024-04077-2
doi:

Substances chimiques

RNA, Bacterial 0

Types de publication

Journal Article

Langues

eng

Sous-ensembles de citation

IM

Pagination

369

Informations de copyright

© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.

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Auteurs

Jignasa H Bera (JH)

Department 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.

Leyon Selvin Raj A (LS)

Department 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.

Hemant Kumar (H)

Department 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.

Nilesh Pandey (N)

Department 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.

Dhara N Patel (DN)

Department 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. dharapatel.cips@charusat.ac.in.
Bapubhai Desaibhai Patel Institute of Paramedical Sciences, Charotar University of Science and Technology, Anand, India. dharapatel.cips@charusat.ac.in.

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Classifications MeSH

questionsmedicales.fr › Acides nucléiques, nucléotides et nucléosides › Acides nucléiques › ARN › ARN bactérien › Effective RNA isolation method for...
questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Cocci à Gram positif › Staphylococcaceae › Staphylococcus › Staphylococcus aureus › Effective RNA isolation method for...
questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Effective RNA isolation method for...
questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Cocci à Gram positif › Staphylococcaceae › Staphylococcus › Staphylococcus epidermidis › Effective RNA isolation method for...
questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Lactobacillales › Enterococcaceae › Enterococcus › Enterococcus faecalis › Effective RNA isolation method for...
questionsmedicales.fr › Bactéries › Bactéries à Gram positif › Bâtonnets à Gram positif › Bâtonnets asporogènes à Gram positif › Bâtonnets asporogènes réguliers à Gram positif › Mycobacteriaceae › Mycobacterium › Mycobactéries non tuberculeuses › Mycobacterium smegmatis › Effective RNA isolation method for...