Utilize este identificador para referenciar este registo: http://hdl.handle.net/10400.12/1722
Título: Evaluation of control transcripts in real-time RT-PCR expression analysis during maritime pine embryogenesis
Autor: Gonçalves, Sónia
Cairney, John
Maroco, João
Oliveira, Maria Margarida
Miguel, Célia M.
Palavras-chave: Housekeeping genes
Maritime pine
Plant embryogenesis
Real-time PCR
Data: 2005
Editora: Springer
Citação: Planta, 222, 556-563
Resumo: In order to determine the suitability of reference or housekeeping genes as internal controls in real-time reverse transcriptase PCR (RT-PCR) assays for quantification of target mRNAs, we studied the levels of expression of four candidate reference genes in maritime pine by real-time RT-PCR. The expression levels obtained for glyceraldehyde-3-phosphate-dehydrogenase, 18S ribosomal RNA, eukaryotic translation initiation factor eIF4AII and ubiquitin in nine stages of embryo development revealed that none of the genes tested proved to be suitable as an internal control. Copy number quantification of the four transcripts showed an average relative variation of seven fold. We propose that the combination of a precise method for RNA quantification, internal controls for monitoring RT reaction and PCR efficiency and a robust external standard curve can guarantee a reliable absolute quantification of mRNA transcripts in real time RT-PCR. This approach may avoid the controversy in the use of housekeeping genes and may assume special significance in tissues undergoing developmental changes.
Peer review: yes
URI: http://hdl.handle.net/10400.12/1722
ISSN: 0032-0935
Aparece nas colecções:BIOL - Artigos em revistas internacionais

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