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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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Circulação/Nível: |
A - 2 |
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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: |
LEADER 02314naa a2200277 a 4500
001 2135836
005 2023-03-10
008 2021 bl uuuu u00u1 u #d
024 7 $a10.1007/s11033-021-06808-w$2DOI
100 1 $aGIGLIOTI, R.
245 $aHow long does the mRNA remains stable in untreated whole bovine blood?$h[electronic resource]
260 $c2021
520 $aBackground 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.
650 $aCattle
650 $aGene expression
650 $aStorage
653 $aFridge
653 $aRNA integrity
653 $aStability
700 1 $aAZEVEDO, B. T.
700 1 $aOLIVEIRA, H. N. DE
700 1 $aKATIKI, L. M.
700 1 $aVERCESI FILHO, A. E.
700 1 $aOLIVEIRA, M. C. de S.
700 1 $aOKINO, C. H.
773 $tMolecular Biology Reports$gv. 49, n. 1, p. 789-795, jan. 2021.
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