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227758

An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus

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Last updated: 28 Dec 2024

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Abstract

Extraction and purification of ribonucleic acid (RNA) from Gram-positive methicillin resistant Staphylococcus aureus (MRSA) is problematic, because the MRSA has a rigid cell wall that contains lipoteichoic acid and peptidoglycan, thus causing difficulty when utilizing the standard methods. For this reason, the aim of the current study was to improve and modify the method of extraction of RNA from MRSA, with good integrity, purity, low cost, and with saved time of extraction. A fast and an inexpensive method involving the use of acid phenol: chloroform (5: 1 [v/v]) at low pH (4.5), with lysostaphin and Triton X-100 for effective isolation of RNA from the MRSA is developed. As a result of this study, yields of this method presented high concentration of RNA (1175.26 ng/ µl/ 3 ml) of bacterial culture broth, with high RNA integration number (RIN). In similar assays such as using; the RNeasy Mini kit, GeneJET RNA purification kit, TRIzol kit and hot phenol: chloroform (1: 1 [v/v]) extraction method, they yielded low concentrations of RNA (92-700 ng/ µl); with lower purity, quantity, and also little integrity, compared to using the current acid phenol chloroform (5: 1 [v/v]) extraction method.  In conclusion, this new method for extraction of RNA from MRSA can be used to save time, cost, and provide high quality of RNA. 

DOI

10.21608/nrmj.2020.84019

Keywords

MRSA, RNA integration number, acid phenol: chloroform, RNA purification

Authors

First Name

Muhanna

Last Name

M. Alshaibani

MiddleName

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Affiliation

School of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia

Email

muhanna@imas.edu.my

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Orcid

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First Name

Noraziah

Last Name

M. Zin

MiddleName

-

Affiliation

School of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia

Email

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Orcid

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First Name

Juriyati

Last Name

Jalil

MiddleName

-

Affiliation

Drug and Herbal Research Centre, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia

Email

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Orcid

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First Name

Anis

Last Name

Rageh Al-Maleki

MiddleName

-

Affiliation

Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia

Email

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Orcid

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First Name

Nik

Last Name

M. Sidik

MiddleName

-

Affiliation

Faculty of Agro-based Industry, Universiti Malaysia Kelantan, 17600 Jeli, Kelantan, Malaysia

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Volume

4

Article Issue

2

Related Issue

32637

Issue Date

2020-04-01

Receive Date

2020-03-24

Publish Date

2020-04-01

Page Start

704

Page End

713

Print ISSN

2537-0286

Online ISSN

2537-0294

Link

https://nrmj.journals.ekb.eg/article_227758.html

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https://nrmj.journals.ekb.eg/service?article_code=227758

Order

227,758

Type

Original Article

Type Code

2,265

Publication Type

Journal

Publication Title

Novel Research in Microbiology Journal

Publication Link

https://nrmj.journals.ekb.eg/

MainTitle

An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus

Details

Type

Article

Created At

23 Jan 2023