Investigating reference genes for quantitative real-time PCR analysis across four chicken tissues
Data de publicació2015-02-11
MetadadesMostra el registre d'unitat complet
Accurate normalization of data is required to correct for different efficiencies and errors during the processing of samples in reverse transcription PCR analysis. The chicken is one of the main livestock species and its genome was one of the first reported and used in large scale transcriptomic analysis.
Despite this, the chicken has not been investigated regarding the identification of reference genes suitable for the quantitative PCR analysis of growth and fattening genes. In this study, five candidate reference genes (B2M, RPL32, SDHA, TBP and YWHAZ) were evaluated to determine the most stable internal reference for quantitative PCR normalization in the two main commercial muscles (pectoralis major (breast) and biceps femoris (thigh)), liver and abdominal fat. Four statistical methods (geNorm, NormFinder, CV and BestKeeper) were used in the evaluation of the most suitable combination of reference genes. Additionally, a comprehensive ranking was established with the RefFinder tool. This analysis identified YWHAZ and TBP as the recommended combination for the analysis of biceps femoris and liver, YWHAZ and RPL32 for pectoralis major and RPL32 and B2M for abdominal fat and across-tissue studies. The final ranking for each tool changed slightly but overall the results, and most particularly the ability to discard the least robust candidates, were consistent between tools. The selection and number of reference genes were validated using SCD, a target gene related to fat metabolism. Overall, the results can be directly used to quantitate target gene expression in different tissues or in validation studies from larger transcriptomic experiments.
És part deGene, 2015, vol. 561, num. 1, p. 82-87
Mostrant elements relacionats per títol, autor i matèria.
Selection of internal control genes for real-time quantitative PCR in ovary and uterus of sows across pregnancy Martinez-Giner, Maria; Noguera Jiménez, José Luis; Balcells, Ingrid; Fernandez-Rodriguez, Amanda; Pena i Subirà, Ramona Natacha (Public Library of Science (PLoS), 2013)Reproductive traits play a key role in pig production in order to reduce costs and increase economic returns. Among others, gene expression analyses represent a useful approach to study genetic mechanisms underlying ...
Genome-Wide Association Study Singles Out SCD and LEPR as the Two Main Loci Influencing Intramuscular Fat Content and Fatty Acid Composition in Duroc Pigs Ros Freixedes, Roger; Gol, Sofia; Pena i Subirà, Ramona Natacha; Tor i Naudí, Marc; Ibañez Escriche, Noelia; Dekkers, Jack C. M.; Estany Illa, Joan (Public Library of Science, 2016)Intramuscular fat (IMF) content and fatty acid composition affect the organoleptic quality and nutritional value of pork. A genome-wide association study was performed on 138 Duroc pigs genotyped with a 60k SNP chip to ...
A functional variant in the Stearoyl-CoA desaturase gene promoter enhances fatty acid desaturation in pork Estany Illa, Joan; Ros Freixedes, Roger; Tor i Naudí, Marc; Pena i Subirà, Ramona Natacha (Public Library of Science (PLoS), 2014)There is growing public concern about reducing saturated fat intake. Stearoyl-CoA desaturase (SCD) is the lipogenic enzyme responsible for the biosynthesis of oleic acid (18:1) by desaturating stearic acid (18:0). Here we ...