An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus
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 thi...
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2020-04-01
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| Series: | Novel Research in Microbiology Journal |
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| Online Access: | https://nrmj.journals.ekb.eg/article_84019_df60ff42739d557492f20be9da68fcb5.pdf |
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| author | Muhanna M. Alshaibani Noraziah M. Zin Juriyati Jalil Anis Rageh Al-Maleki Nik M. Sidik |
| author_facet | Muhanna M. Alshaibani Noraziah M. Zin Juriyati Jalil Anis Rageh Al-Maleki Nik M. Sidik |
| author_sort | Muhanna M. Alshaibani |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-3ce6864be193477fae12b36a443484e6 |
| institution | Kabale University |
| issn | 2537-0286 2537-0294 |
| language | English |
| publishDate | 2020-04-01 |
| publisher | ResearchersLinks, Ltd |
| record_format | Article |
| series | Novel Research in Microbiology Journal |
| spelling | doaj-art-3ce6864be193477fae12b36a443484e62025-08-20T03:55:33ZengResearchersLinks, LtdNovel Research in Microbiology Journal2537-02862537-02942020-04-014270471310.21608/nrmj.2020.84019An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureusMuhanna M. Alshaibani0Noraziah M. Zin1Juriyati Jalil2Anis Rageh Al-Maleki3Nik M. Sidik4School of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, MalaysiaSchool of Diagnostic and Applied Health Sciences, Faculty of Health Sciences, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, MalaysiaDrug and Herbal Research Centre, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300 Kuala Lumpur, MalaysiaDepartment of Medical Microbiology, Faculty of Medicine, University of Malaya, Kuala Lumpur, 50603, Malaysia; Department of Medical Microbiology, Faculty of Medicine and Health Sciences, Sana’a University, Sana’a, YemenFaculty of Agro-based Industry, Universiti Malaysia Kelantan, 17600 Jeli, Kelantan, MalaysiaExtraction 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.https://nrmj.journals.ekb.eg/article_84019_df60ff42739d557492f20be9da68fcb5.pdfmrsarna integration numberacid phenol: chloroformrna purification |
| spellingShingle | Muhanna M. Alshaibani Noraziah M. Zin Juriyati Jalil Anis Rageh Al-Maleki Nik M. Sidik An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus Novel Research in Microbiology Journal mrsa rna integration number acid phenol: chloroform rna purification |
| title | An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus |
| title_full | An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus |
| title_fullStr | An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus |
| title_full_unstemmed | An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus |
| title_short | An improved procedure for the isolation of Ribonucleic acid from methicillin-resistant Staphylococcus aureus |
| title_sort | improved procedure for the isolation of ribonucleic acid from methicillin resistant staphylococcus aureus |
| topic | mrsa rna integration number acid phenol: chloroform rna purification |
| url | https://nrmj.journals.ekb.eg/article_84019_df60ff42739d557492f20be9da68fcb5.pdf |
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