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 | Acesso ao texto completo restrito à biblioteca da Embrapa Pecuária Sudeste. Para informações adicionais entre em contato com cppse.biblioteca@embrapa.br. |
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Registro Completo |
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Biblioteca(s): |
Embrapa Pecuária Sudeste. |
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Data corrente: |
05/11/2021 |
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Data da última atualização: |
10/03/2023 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Autoria: |
GIGLIOTI, R.; AZEVEDO, B. T.; OLIVEIRA, H. N. DE; KATIKI, L. M.; VERCESI FILHO, A. E.; OLIVEIRA, M. C. de S.; OKINO, C. H. |
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Afiliação: |
RODRIGO GIGLIOTI, IZ; BIANCA TAINÁ AZEVEDO, IZ; HENRIQUE NUNES DE OLIVEIRA, UNESP; LUCIANA MORITA KATIKI, IZ; ANIBAL EUGÊNIO VERCESI FILHO, IZ; MARCIA CRISTINA DE SENA OLIVEIRA, CPPSE; CINTIA HIROMI OKINO, CPPSE. |
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Título: |
How long does the mRNA remains stable in untreated whole bovine blood? |
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Ano de publicação: |
2021 |
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Fonte/Imprenta: |
Molecular Biology Reports, v. 49, n. 1, p. 789-795, jan. 2021. |
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DOI: |
10.1007/s11033-021-06808-w |
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Idioma: |
Inglês
Português |
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Conteúdo: |
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 specifc 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 diferent 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 diferent 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 success-fully and accurately used for gene expression studies. MenosBackground 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 specifc 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 diferent 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 diferent 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 success-fully and accurately used ... Mostrar Tudo |
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Palavras-Chave: |
Fridge; RNA integrity; Stability. |
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Thesaurus Nal: |
Cattle; Gene expression; Storage. |
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Categoria do assunto: |
G Melhoramento Genético
K Ciência Florestal e Produtos de Origem Vegetal |
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Marc: |
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Registro original: |
Embrapa Pecuária Sudeste (CPPSE) |
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| 3. |  | VIEIRA, A. L. M.; ZAMBONIM, F. M.; LISBOA, A. C.; OLIVEIRA, N. M.; FREITAS, A. F. N.; RESENDE, A. S.; CAMPELLO, E. F. C. Estrutura de Paisagem e distribuição de pequenos mamíferos: Importância de um Corredor agro florestal em uma área de mata Atlântica In: SEMANA CIENTÍFICA JOHANNA DOBEREINER, 9., 19 a 23 de outubro de 2009. Seropédica: Embrapa Agrobiologia, 2009 01 01| Tipo: Folder/Folheto/Cartilha |
| Biblioteca(s): Embrapa Agrobiologia. |
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| 4. | | LISBOA, A. C.; MELO JÚNIOR, C. J. A. H. de; TAVARES, F. P. A.; ALMEIDA, R. B. de; MELO, L. A. de; MAGISTRALI, I. C. Crescimento e qualidade de mudas de Handroanthus heptaphyllus em substrato com esterco bovino. Pesquisa Florestal Brasileira, Colombo, v. 38, e201701485, 2018. 6 p. Nota científica.| Biblioteca(s): Embrapa Florestas. |
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| 5. | | BARRETO, L. M. G.; MEDEIROS, A. N. de; BATISTA, A. M. V.; FURTADO, D. A.; ARAUJO, G. G. L. de; LISBOA, A. C. C.; PAULO, J. L. de A.; SOUZA, C. M. S. de. Comportamento ingestivo de caprinos das raças Moxotó e Canindé em confinamento recebendo dois níveis de energia na dieta. Revista Brasileira de Zootecnia, v. 40, n. 4, p. 834-842, 2011.| Tipo: Artigo em Periódico Indexado | Circulação/Nível: B - 1 |
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