Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus
Biofilm formed by Staphylococcus aureus is considered an important virulence trait in the pathogenesis of infections associated with implantable medical devices. Gene expression analyses are important strategies for determining the mechanisms involved in production and regulation of biofilm. Obtaini...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Associação Brasileira de Divulgação Científica
2015-12-01
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| Series: | Brazilian Journal of Medical and Biological Research |
| Subjects: | |
| Online Access: | http://www.scielo.br/pdf/bjmbr/v48n12/1414-431X-bjmbr-1414-431X20154734.pdf |
| _version_ | 1798013898175021056 |
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| author | C. O. Beltrame M. F. Côrtes P. T. Bandeira A. M. S. Figueiredo |
| author_facet | C. O. Beltrame M. F. Côrtes P. T. Bandeira A. M. S. Figueiredo |
| author_sort | C. O. Beltrame |
| collection | DOAJ |
| description | Biofilm formed by Staphylococcus aureus is considered an important virulence trait in the pathogenesis of infections associated with implantable medical devices. Gene expression analyses are important strategies for determining the mechanisms involved in production and regulation of biofilm. Obtaining intact RNA preparations is the first and most critical step for these studies. In this article, we describe an optimized protocol for obtaining total RNA from sessile cells of S. aureus using the RNeasy Mini Kit. This method essentially consists of a few steps, as follows: 1) addition of acetone-ethanol to sessile cells, 2) lysis with lysostaphin at 37°C/10 min, 3) vigorous mixing, 4) three cycles of freezing and thawing, and 5) purification of the lysate in the RNeasy column. This simple pre-kit procedure yields high-quality total RNA from planktonic and sessile cells of S. aureus. |
| first_indexed | 2024-04-11T15:09:58Z |
| format | Article |
| id | doaj.art-e82e58ff84364aac80311c598198475d |
| institution | Directory Open Access Journal |
| issn | 1414-431X |
| language | English |
| last_indexed | 2024-04-11T15:09:58Z |
| publishDate | 2015-12-01 |
| publisher | Associação Brasileira de Divulgação Científica |
| record_format | Article |
| series | Brazilian Journal of Medical and Biological Research |
| spelling | doaj.art-e82e58ff84364aac80311c598198475d2022-12-22T04:16:42ZengAssociação Brasileira de Divulgação CientíficaBrazilian Journal of Medical and Biological Research1414-431X2015-12-0148121071107610.1590/1414-431x20154734Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureusC. O. BeltrameM. F. CôrtesP. T. BandeiraA. M. S. FigueiredoBiofilm formed by Staphylococcus aureus is considered an important virulence trait in the pathogenesis of infections associated with implantable medical devices. Gene expression analyses are important strategies for determining the mechanisms involved in production and regulation of biofilm. Obtaining intact RNA preparations is the first and most critical step for these studies. In this article, we describe an optimized protocol for obtaining total RNA from sessile cells of S. aureus using the RNeasy Mini Kit. This method essentially consists of a few steps, as follows: 1) addition of acetone-ethanol to sessile cells, 2) lysis with lysostaphin at 37°C/10 min, 3) vigorous mixing, 4) three cycles of freezing and thawing, and 5) purification of the lysate in the RNeasy column. This simple pre-kit procedure yields high-quality total RNA from planktonic and sessile cells of S. aureus.http://www.scielo.br/pdf/bjmbr/v48n12/1414-431X-bjmbr-1414-431X20154734.pdfRNA preparationMRSABiofilmSessile cellsStaphylococcus aureus |
| spellingShingle | C. O. Beltrame M. F. Côrtes P. T. Bandeira A. M. S. Figueiredo Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus Brazilian Journal of Medical and Biological Research RNA preparation MRSA Biofilm Sessile cells Staphylococcus aureus |
| title | Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus |
| title_full | Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus |
| title_fullStr | Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus |
| title_full_unstemmed | Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus |
| title_short | Optimization of the RNeasy Mini Kit to obtain high-quality total RNA from sessile cells of Staphylococcus aureus |
| title_sort | optimization of the rneasy mini kit to obtain high quality total rna from sessile cells of staphylococcus aureus |
| topic | RNA preparation MRSA Biofilm Sessile cells Staphylococcus aureus |
| url | http://www.scielo.br/pdf/bjmbr/v48n12/1414-431X-bjmbr-1414-431X20154734.pdf |
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