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Real time PCR and importance of housekeepings genes for normalization and quantification of mRNA expression in different tissues
Rebouças, Emanuela de Lima; Costa, José Jackson do Nascimento; Passos, Maria Juliane; Passos, José Renato de Sousa; Hurk, Robert van den; Silva, José Roberto Viana.
  • Rebouças, Emanuela de Lima; Universidade Federal do Ceara. Nucleo de Biotecnologia de Sobral. Sobral. BR
  • Costa, José Jackson do Nascimento; Universidade Federal do Ceara. Nucleo de Biotecnologia de Sobral. Sobral. BR
  • Passos, Maria Juliane; Universidade Federal do Ceara. Nucleo de Biotecnologia de Sobral. Sobral. BR
  • Passos, José Renato de Sousa; Universidade Federal do Ceara. Nucleo de Biotecnologia de Sobral. Sobral. BR
  • Hurk, Robert van den; Universidade Federal do Ceara. Nucleo de Biotecnologia de Sobral. Sobral. BR
  • Silva, José Roberto Viana; Universidade Federal do Ceara. Nucleo de Biotecnologia de Sobral. Sobral. BR
Braz. arch. biol. technol ; 56(1): 143-154, Jan.-Feb. 2013. tab
Artículo en Inglés | LILACS | ID: lil-670293
ABSTRACT
The aim of this review was to evaluate the importance of the real-time PCR (qRT-PCR) as a technique for mRNA expression analysis in different tissues. Real-time PCR is widely used for quantification of mRNA levels and is a fundamental tool for basic research, molecular medicine and biotechnology.Genes of references are expressed in a wide variety of tissues and cells with minimal variations in their expression levels, and thus are used to normalize data of mRNA quantification. Software programs, such as geNorm, BestKeeper and NormFinder, have been developed to perform the normalization of data, which help to choose the most stable reference gene. Several genes, such as GAPDH, β-actin, β-tubulin, PGK, UBQ, RPL-19 and 18S rRNA have been suggested as standards in PCR studies, but these genes can have variation in their expression in different tissues, reinforcing the idea that there is no ideal reference gene.


Texto completo: Disponible Índice: LILACS (Américas) Idioma: Inglés Revista: Braz. arch. biol. technol Asunto de la revista: Biologia Año: 2013 Tipo del documento: Artículo País de afiliación: Brasil Institución/País de afiliación: Universidade Federal do Ceara/BR

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Texto completo: Disponible Índice: LILACS (Américas) Idioma: Inglés Revista: Braz. arch. biol. technol Asunto de la revista: Biologia Año: 2013 Tipo del documento: Artículo País de afiliación: Brasil Institución/País de afiliación: Universidade Federal do Ceara/BR