Stable reference genes for expression studies in breast muscle of normal and white striping-affected chickens.
Caroline Michele Marinho Marciano, Adriana Mércia Guaratini Ibelli, Jane de Oliveira Peixoto, Igor Ricardo Savoldi, Kamilla Bleil do Carmo, Lana Teixeira Fernandes, Mônica Corrêa Ledur
Author Information
Caroline Michele Marinho Marciano: Programa de Pós-Graduação em Zootecnia, Universidade do Estado de Santa Catarina, UDESC-Oeste, Rua Beloni Trombeta Zanin 680E, Chapecó, Santa Catarina, 89815-630, Brazil. ORCID
Adriana Mércia Guaratini Ibelli: Embrapa Suínos e Aves, Rodovia BR-153, Km 110, Distrito de Tamanduá, Caixa Postal: 321, Concórdia, Santa Catarina, 89715-899, Brazil. ORCID
Jane de Oliveira Peixoto: Embrapa Suínos e Aves, Rodovia BR-153, Km 110, Distrito de Tamanduá, Caixa Postal: 321, Concórdia, Santa Catarina, 89715-899, Brazil. ORCID
Igor Ricardo Savoldi: Programa de Pós-Graduação em Zootecnia, Universidade do Estado de Santa Catarina, UDESC-Oeste, Rua Beloni Trombeta Zanin 680E, Chapecó, Santa Catarina, 89815-630, Brazil. ORCID
Kamilla Bleil do Carmo: Universidade do Contestado, Rua Victor Sopelsa, 3000, Concórdia, Santa Catarina, 89711-330, Brazil. ORCID
Lana Teixeira Fernandes: Centro de Diagnóstico de Sanidade Animal, Rodovia BR-153, Km 110, Distrito de Tamanduá, Caixa Postal: 321, Concórdia, Santa Catarina, 89715-899, Brazil. ORCID
Mônica Corrêa Ledur: Programa de Pós-Graduação em Zootecnia, Universidade do Estado de Santa Catarina, UDESC-Oeste, Rua Beloni Trombeta Zanin 680E, Chapecó, Santa Catarina, 89815-630, Brazil. monica.ledur@embrapa.br. ORCID
The normalization with proper reference genes is a crucial step to obtain accurate mRNA expression levels in quantitative PCR (qPCR) studies. Therefore, in this study, 10 reference candidate genes were evaluated to determine their stability in normal pectoralis major muscle of broilers and those counterparts affected with White Striping (WS) myopathy at 42 days age. Four different tools were used for ranking the most stable genes: GeNorm, NormFinder, BestKeeper and Comparative Ct (ΔCt), and a general ranking was performed using the RankAggreg tool to select the best reference genes among all tools. From the 10 genes evaluated in the breast muscle of broilers, 8 were amplified. Most of the algorithms/tools indicated the same two genes, RPL30 and RPL5, as the most stable in the broilers breast muscle. In addition, there was agreement among the tools for the least stable genes: MRPS27, GAPDH and RPLP1 in the broilers breast muscle. Therefore, it is interesting to note that even with different tools for evaluating gene expression, there was consensus on the most and least stable genes. These results indicate that the Ribosomal protein L30 (RPL30) and Ribosomal protein L5 (RPL5) can be recommended for accurate normalization in qPCR studies with chicken pectoralis major muscle affected with White Striping and other myopathies.
