How long does the mRNA remains stable in untreated whole bovine blood?
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1007/s11033-021-06808-w http://hdl.handle.net/11449/222638 |
Resumo: | Background: High quality and quantity of messenger RNA (mRNA) are required for accuracy of gene expression studies and other RNA-based downstream applications. Since RNA is considered a labile macromolecular prone to degradation, which may result in falsely altered gene expression patterns, several commercial stabilizing reagents have been developed aiming to keep RNA stable for long period. However, for studies involving large number of experimental samples, the high costs related to these specific reagents may constitute a barrier. Methods and results: In this context the present study was designed aiming to evaluate the stability of mRNA in whole bovine blood collected in EDTA tubes during storage at common fridge (4 °C). Whole blood samples were collected from six Holstein calves and submitted to RNA extraction in each different interval: immediately after blood sampling (< 2 h), at 1-day post-sampling (dps), 2 dps, 3 dps, 7 dps and 14dps intervals. RNA integrity and purity were evaluated, and RT-qPCR assays were run using seven different genes (B2M, ACTB, PPIA, GAPDH, YWHAZ, CD4 and IFN-γ) aiming to evaluate the presence of altered gene transcription during storage. All extracted RNA samples presented high purity, while optimal integrity and unaltered gene expression were observed in whole experimental group up to 3 days of storage. Conclusion: Bovine blood RNA remained stable in K3EDTA tubes for 3 days stored at common fridge and can be successfully and accurately used for gene expression studies. |
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How long does the mRNA remains stable in untreated whole bovine blood?CattleFridgeGene expressionRNA integrityStabilityStorageBackground: High quality and quantity of messenger RNA (mRNA) are required for accuracy of gene expression studies and other RNA-based downstream applications. Since RNA is considered a labile macromolecular prone to degradation, which may result in falsely altered gene expression patterns, several commercial stabilizing reagents have been developed aiming to keep RNA stable for long period. However, for studies involving large number of experimental samples, the high costs related to these specific reagents may constitute a barrier. Methods and results: In this context the present study was designed aiming to evaluate the stability of mRNA in whole bovine blood collected in EDTA tubes during storage at common fridge (4 °C). Whole blood samples were collected from six Holstein calves and submitted to RNA extraction in each different interval: immediately after blood sampling (< 2 h), at 1-day post-sampling (dps), 2 dps, 3 dps, 7 dps and 14dps intervals. RNA integrity and purity were evaluated, and RT-qPCR assays were run using seven different genes (B2M, ACTB, PPIA, GAPDH, YWHAZ, CD4 and IFN-γ) aiming to evaluate the presence of altered gene transcription during storage. All extracted RNA samples presented high purity, while optimal integrity and unaltered gene expression were observed in whole experimental group up to 3 days of storage. Conclusion: Bovine blood RNA remained stable in K3EDTA tubes for 3 days stored at common fridge and can be successfully and accurately used for gene expression studies.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Instituto de Zootecnia, Rua Heitor Penteado, n. 56,São PauloUniversidade Estadual Júlio de Mesquita Filho Unesp, São PauloEmbrapa Pecuária Sudeste, São PauloUniversidade Estadual Júlio de Mesquita Filho Unesp, São PauloFAPESP: 2016/07216-7FAPESP: 2019/22675-6Instituto de ZootecniaUniversidade Estadual Paulista (UNESP)Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)Giglioti, RodrigoAzevedo, Bianca Taináde Oliveira, Henrique Nunes [UNESP]Katiki, Luciana MoritaFilho, Anibal Eugênio Verceside Sena Oliveira, Márcia CristinaOkino, Cintia Hiromi2022-04-28T19:45:55Z2022-04-28T19:45:55Z2022-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article789-795http://dx.doi.org/10.1007/s11033-021-06808-wMolecular Biology Reports, v. 49, n. 1, p. 789-795, 2022.1573-49780301-4851http://hdl.handle.net/11449/22263810.1007/s11033-021-06808-w2-s2.0-85117029180Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengMolecular Biology Reportsinfo:eu-repo/semantics/openAccess2022-04-28T19:45:55Zoai:repositorio.unesp.br:11449/222638Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T21:08:20.696139Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
How long does the mRNA remains stable in untreated whole bovine blood? |
title |
How long does the mRNA remains stable in untreated whole bovine blood? |
spellingShingle |
How long does the mRNA remains stable in untreated whole bovine blood? Giglioti, Rodrigo Cattle Fridge Gene expression RNA integrity Stability Storage |
title_short |
How long does the mRNA remains stable in untreated whole bovine blood? |
title_full |
How long does the mRNA remains stable in untreated whole bovine blood? |
title_fullStr |
How long does the mRNA remains stable in untreated whole bovine blood? |
title_full_unstemmed |
How long does the mRNA remains stable in untreated whole bovine blood? |
title_sort |
How long does the mRNA remains stable in untreated whole bovine blood? |
author |
Giglioti, Rodrigo |
author_facet |
Giglioti, Rodrigo Azevedo, Bianca Tainá de Oliveira, Henrique Nunes [UNESP] Katiki, Luciana Morita Filho, Anibal Eugênio Vercesi de Sena Oliveira, Márcia Cristina Okino, Cintia Hiromi |
author_role |
author |
author2 |
Azevedo, Bianca Tainá de Oliveira, Henrique Nunes [UNESP] Katiki, Luciana Morita Filho, Anibal Eugênio Vercesi de Sena Oliveira, Márcia Cristina Okino, Cintia Hiromi |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Instituto de Zootecnia Universidade Estadual Paulista (UNESP) Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) |
dc.contributor.author.fl_str_mv |
Giglioti, Rodrigo Azevedo, Bianca Tainá de Oliveira, Henrique Nunes [UNESP] Katiki, Luciana Morita Filho, Anibal Eugênio Vercesi de Sena Oliveira, Márcia Cristina Okino, Cintia Hiromi |
dc.subject.por.fl_str_mv |
Cattle Fridge Gene expression RNA integrity Stability Storage |
topic |
Cattle Fridge Gene expression RNA integrity Stability Storage |
description |
Background: High quality and quantity of messenger RNA (mRNA) are required for accuracy of gene expression studies and other RNA-based downstream applications. Since RNA is considered a labile macromolecular prone to degradation, which may result in falsely altered gene expression patterns, several commercial stabilizing reagents have been developed aiming to keep RNA stable for long period. However, for studies involving large number of experimental samples, the high costs related to these specific reagents may constitute a barrier. Methods and results: In this context the present study was designed aiming to evaluate the stability of mRNA in whole bovine blood collected in EDTA tubes during storage at common fridge (4 °C). Whole blood samples were collected from six Holstein calves and submitted to RNA extraction in each different interval: immediately after blood sampling (< 2 h), at 1-day post-sampling (dps), 2 dps, 3 dps, 7 dps and 14dps intervals. RNA integrity and purity were evaluated, and RT-qPCR assays were run using seven different genes (B2M, ACTB, PPIA, GAPDH, YWHAZ, CD4 and IFN-γ) aiming to evaluate the presence of altered gene transcription during storage. All extracted RNA samples presented high purity, while optimal integrity and unaltered gene expression were observed in whole experimental group up to 3 days of storage. Conclusion: Bovine blood RNA remained stable in K3EDTA tubes for 3 days stored at common fridge and can be successfully and accurately used for gene expression studies. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-04-28T19:45:55Z 2022-04-28T19:45:55Z 2022-01-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1007/s11033-021-06808-w Molecular Biology Reports, v. 49, n. 1, p. 789-795, 2022. 1573-4978 0301-4851 http://hdl.handle.net/11449/222638 10.1007/s11033-021-06808-w 2-s2.0-85117029180 |
url |
http://dx.doi.org/10.1007/s11033-021-06808-w http://hdl.handle.net/11449/222638 |
identifier_str_mv |
Molecular Biology Reports, v. 49, n. 1, p. 789-795, 2022. 1573-4978 0301-4851 10.1007/s11033-021-06808-w 2-s2.0-85117029180 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Molecular Biology Reports |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
789-795 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1808129289845997568 |