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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: |
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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Registro original: |
Embrapa Pecuária Sudeste (CPPSE) |
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Biblioteca |
ID |
Origem |
Tipo/Formato |
Classificação |
Cutter |
Registro |
Volume |
Status |
URL |
Voltar
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Registro Completo
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Biblioteca(s): |
Embrapa Milho e Sorgo. |
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Data corrente: |
10/03/2009 |
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Data da última atualização: |
23/05/2018 |
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Tipo da produção científica: |
Artigo em Periódico Indexado |
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Circulação/Nível: |
Nacional - B |
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Autoria: |
MARTINS, A. O.; CAMPOSTRINI, E.; MAGALHAES, P. C.; GUIMARAES, L. J. M.; DURAES, F. O. M.; MARRIEL, I. E.; NETTO, A. T. |
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Afiliação: |
Amanda Oliveira Martins, UENF; Eliemar Campostrini, UENF; PAULO CESAR MAGALHAES, CNPMS; LAURO JOSE MOREIRA GUIMARAES, CNPMS; Frederico Ozanan Machado Durães, Embrapa Milho e Sorgo; IVANILDO EVODIO MARRIEL, CNPMS; Alena Torres Netto, UENF. |
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Título: |
Nitrogen-use efficiency of maize genotypes in contrasting environments. |
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Ano de publicação: |
2008 |
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Fonte/Imprenta: |
Crop Breeding and Applied Biotechnology, Londrina, v. 8, n. 4, p. 291-298, 2008. |
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Idioma: |
Inglês |
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Conteúdo: |
The purpose ofthis study was to evaluate the nitrogen-use efficiency of 15 corn hybrids in different environments. The experiment was conducted at the research station Embrapa Milho e Sorgo, in environments with low (12 kg ha-l) and high (120 kg ha-J) nitrogen fertilization, in a randomized block design with three replications. The chlorophyll content, leaf nitrogen and nitrate reductase activity in the flag leaf were determined at flowering and the grain yield was evaluated at harvest. The chlorophyll content proved to be sensitive to nitrogen availability, although without discriminating genotypic differences efficiently. To use nitrate reductasefor this purposeJurther studies are needed on the reliability ofthis biochemical variable to diagnose genotypes efficient in N use precociously. The hybrid L2xL3 was considered promising for maize breeding programs aimed at gene introgression related to N-use efficiency. |
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Palavras-Chave: |
Chlorophyll content; Leafnitrogen; Yield. |
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Thesagro: |
Zea Mays. |
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Thesaurus NAL: |
nitrogen. |
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Categoria do assunto: |
S Ciências Biológicas |
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URL: |
https://ainfo.cnptia.embrapa.br/digital/bitstream/item/66413/1/Nitrogen-use.pdf
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Marc: |
LEADER 01657naa a2200253 a 4500
001 1491822
005 2018-05-23
008 2008 bl uuuu u00u1 u #d
100 1 $aMARTINS, A. O.
245 $aNitrogen-use efficiency of maize genotypes in contrasting environments.$h[electronic resource]
260 $c2008
520 $aThe purpose ofthis study was to evaluate the nitrogen-use efficiency of 15 corn hybrids in different environments. The experiment was conducted at the research station Embrapa Milho e Sorgo, in environments with low (12 kg ha-l) and high (120 kg ha-J) nitrogen fertilization, in a randomized block design with three replications. The chlorophyll content, leaf nitrogen and nitrate reductase activity in the flag leaf were determined at flowering and the grain yield was evaluated at harvest. The chlorophyll content proved to be sensitive to nitrogen availability, although without discriminating genotypic differences efficiently. To use nitrate reductasefor this purposeJurther studies are needed on the reliability ofthis biochemical variable to diagnose genotypes efficient in N use precociously. The hybrid L2xL3 was considered promising for maize breeding programs aimed at gene introgression related to N-use efficiency.
650 $anitrogen
650 $aZea Mays
653 $aChlorophyll content
653 $aLeafnitrogen
653 $aYield
700 1 $aCAMPOSTRINI, E.
700 1 $aMAGALHAES, P. C.
700 1 $aGUIMARAES, L. J. M.
700 1 $aDURAES, F. O. M.
700 1 $aMARRIEL, I. E.
700 1 $aNETTO, A. T.
773 $tCrop Breeding and Applied Biotechnology, Londrina$gv. 8, n. 4, p. 291-298, 2008.
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Registro original: |
Embrapa Milho e Sorgo (CNPMS) |
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