An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus | ||||
Novel Research in Microbiology Journal | ||||
Volume 4, Issue 2, March and April 2020, Page 704-713 PDF (261.53 K) | ||||
Document Type: Original Article | ||||
DOI: 10.21608/nrmj.2020.84019 | ||||
View on SCiNiTO | ||||
Authors | ||||
Muhanna M. Alshaibani 1; Noraziah M. Zin1; Juriyati Jalil2; Anis Rageh Al-Maleki3; Nik M. Sidik4 | ||||
1School of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia | ||||
2Drug and Herbal Research Centre, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, Malaysia | ||||
3Department of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia | ||||
4Faculty of Agro-based Industry, Universiti Malaysia Kelantan, 17600 Jeli, Kelantan, Malaysia | ||||
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. | ||||
Keywords | ||||
MRSA; RNA integration number; acid phenol: chloroform; RNA purification | ||||
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