Reference gene selection for reverse transcription quantitative polymerase chain reaction in chicken hypothalamus under different feeding status.
J Anim Physiol Anim Nutr (Berl)
; 102(1): 286-296, 2018 Feb.
Article
en En
| MEDLINE
| ID: mdl-28294410
ABSTRACT
This study was designed to investigate the stability of 10 candidate reference genes, namely ACTB, B2M, GAPDH, HMBS, LBR, POLR2B, RN18S, RPS17, TBP, and YWHAZ for the normalization of gene expression data obtained by quantitative real-time polymerase chain reaction (qPCR) in studies related to feed intake of chicken. Samples were isolated from hypothalamus under three different nutritional status (ad libitum, fasted for 24 hr, fasted for 24 hr then refed for 2 hr). Five different algorithms were applied for the analysis of reference gene stability BestKeeper, geNorm, NormFinder, the comparative ΔCt method, and a novel approach using multivariate linear mixed-effects modelling for stable reference gene selection. TBP and POLR2B were identified as the two most suitable and B2M and RN18S as the two least stable reference genes for normalization. Despite our review, the current literature showing that RN18S is one of the most commonly used reference gene in chicken gene expression studies, its applicability for normalization should be evaluated before each qPCR experiment.
Palabras clave
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Pollos
/
Regulación de la Expresión Génica
/
Reacción en Cadena de la Polimerasa de Transcriptasa Inversa
/
Privación de Alimentos
/
Hipotálamo
Límite:
Animals
Idioma:
En
Revista:
J Anim Physiol Anim Nutr (Berl)
Asunto de la revista:
CIENCIAS DA NUTRICAO
/
FISIOLOGIA
/
MEDICINA VETERINARIA
Año:
2018
Tipo del documento:
Article
País de afiliación:
Hungria