Modified Spin Column-Based RNA Extraction Methods of Staphylococcus aureus using PureLink® RNA Mini Kit and Basic Laboratory Instrument
Article Sidebar
Main Article Content
https://orcid.org/0000-0001-9497-2465
Abstract
RNA extraction is an important process before gene expression assessment at the transcriptomic level. RNA is a sensitive material to environmental factors such as temperature and contaminants, so the RNA extraction process generally requires sophisticated and expensive laboratory instruments. In this study, we extract RNA from Staphylococcus aureus bacteria using the PureLink® RNA Mini Kit. The instruments used in this study are basic instruments such as a hand homogenizer and non-thermal centrifuge. The results of RNA extraction were visualized using agarose gel electrophoresis. These results indicate that bacterial RNA extraction can be performed using the PureLink® RNA Mini Kit even with inexpensive basic laboratory instruments.
Downloads
Article Details
Copyright (c) 2021 Muhammad Taufiq Hidayat, Endry Nugroho Prasetyo
This work is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License.
References
Wang H, Han Y. Chapter 4 Extraction of total RNA from Yersinia pestis. Yersinia pestis protoc., Singapore: Springer; 2018, p. 29–34.
Rodrıguez A, Duyvejonck H, Belleghem JD Van, Gryp T, Simaey L Van, Vermeulen S, et al. Comparison of procedures for RNA-extraction from peripheral blood mononuclear cells. PlosOne 2020;15:1–17. https://doi.org/10.1371/journal.pone.0229423.
Ahmad J, Baig MA, Ali AA, Al-Huqail A, Ibrahim MM, Qureshi MI. Comparative assessment of four RNA extraction methods and modification to obtain high-quality RNA from Parthenium hysterophorus leaf. 3 Biotech 2017;7:373. https://doi.org/10.1007/s13205-017-1003-3.
Bezerra D da S, Neves BG, Guedes SF de F, Regis WFM, Stipp RN, Rodrigues LKA. Extraction and purification of RNA from human carious dentine : an approach to enable bacterial gene expression studies. J Heal Biol Sci 2019;7:145–51. https://doi.org/10.12662/2317-3076jhbs.v7i2.2562.p145-151.2019.
Ambion. PureLink ® RNA Mini Kit. Carlsbad: Life Technologies Corporation; 2012.
Zavala-Alvarado, Crispin, Benaroudj N. The Single-Step Method of RNA Purification Applied to Leptospira. Leptospira spp, New York: Humana; 2020, p. 41–51.
Kitichalermkiat A, Katsuki M, Sato J, Sonoda T, Masuda Y, Honjoh K ichi, et al. Effect of epigallocatechin gallate on gene expression of Staphylococcus aureus. J Glob Antimicrob Resist 2020;22:854–9. https://doi.org/10.1016/j.jgar.2020.06.006.
Liu Y, Breukink E. The membrane steps of bacterial cell wall synthesis as antibiotic targets. Antibiotics 2016;5:1–22. https://doi.org/10.3390/antibiotics5030028.
Shi R, Lewis RS, Panthee DR. Filter paper-based spin column method for cost-efficient DNA or RNA purification. PlosOne 2018;13:1–14.
Rodríguez ML, Rosa AC, Jewtuchowicz V. RNA extraction from the yeast Candida parapsilosis sensu stricto using two commercially available silica column-based purification methods. J Microbiol Mod Tech 2018;3:201.
Kher MN, Sheth NR, Bhatt VD. In vitro antibacterial evaluation of terminalia chebula as an alternative of antibiotics against bovine subclinical mastitis. Anim Biotechnol 2019;30:151–8. https://doi.org/10.1080/10495398.2018.1451752.
Nadiya F, Anjali N, Gangaprasad A, Krishnannair K. High-quality RNA extraction from small cardamom tissues rich in polysaccharides and polyphenols. Anal Biochem 2015;485:25–7. https://doi.org/10.1016/j.ab.2015.05.017.
Bhat AI, Rao GP. Agarose gel electrophoresis for nucleic acids. Charact. Plant Viruses. Springer Protoc. Handbooks, New York: Humana; 2020.
Hu WP. Improve sample preparation process for miRNA isolation from the culture cells by using silica fiber membrane. Sci Rep 2020;10:1–9. https://doi.org/10.1038/s41598-020-78202-8.
Yang F, Wang G, Xu W, Hong N. A rapid silica spin column-based method of RNA extraction from fruit trees for RT-PCR detection of viruses. J Virol Methods 2017;247:61–7. https://doi.org/10.1016/j.jviromet.2017.05.020.
Xu R, Shieh YC, Stewart DS. Comparison of RNA extraction kits for the purification and detection of an enteric virus surrogate on green onions via RT-PCR. J Virol Methods 2017;239:61–8. https://doi.org/10.1016/j.jviromet.2016.10.016.
Hay M, Li YM, Ma Y. RNA extraction of Escherichia coli grown in lysogeny broth for use in RT-qPCR. JEMI Methods 2017;1:1–6.
Wever J De, Tulkens D, Verwaeren J, Everaert H, Rottiers H, Dewettinck K, et al. A combined rna preservation and extraction protocol for gene expression studies in cacao beans. Front Plant Sci 2020;11:992. https://doi.org/10.3389/fpls.2020.00992.
Breitler J, Campa C, Georget F, Bertrand B, Etienne H. A single-step method for RNA isolation from tropical crops in the field. Nat Publ Gr 2016;6:1–6. https://doi.org/10.1038/srep38368.
Abdallah NMA, Zaki AM, Abdel-salam SA. Stability of MERS-CoV RNA on spin columns of RNA extraction kit at room temperature. Diagnostic Microbiol Infect Dis 2020;98:115182. https://doi.org/10.1016/j.diagmicrobio.2020.115182.
