Selection of reference genes for gene expression studies related to intramuscular fat deposition in Capra hircus skeletal muscle

PloS One
Wuzheng ZhuYong Wang

Abstract

The identification of suitable reference genes is critical for obtaining reliable results from gene expression studies using quantitative real-time PCR (qPCR) because the expression of reference genes may vary considerably under different experimental conditions. In most cases, however, commonly used reference genes are employed in data normalization without proper validation, which may lead to incorrect data interpretation. Here, we aim to select a set of optimal reference genes for the accurate normalization of gene expression associated with intramuscular fat (IMF) deposition during development. In the present study, eight reference genes (PPIB, HMBS, RPLP0, B2M, YWHAZ, 18S, GAPDH and ACTB) were evaluated by three different algorithms (geNorm, NormFinder and BestKeeper) in two types of muscle tissues (longissimus dorsi muscle and biceps femoris muscle) across different developmental stages. All three algorithms gave similar results. PPIB and HMBS were identified as the most stable reference genes, while the commonly used reference genes 18S and GAPDH were the most variably expressed, with expression varying dramatically across different developmental stages. Furthermore, to reveal the crucial role of appropriate reference ge...Continue Reading

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Citations

Mar 1, 2017·Clinica Chimica Acta; International Journal of Clinical Chemistry·Yu JiangYibin Guo
May 24, 2016·Scientific Reports·Prue M Pereira-FantiniDavid G Tingay
Aug 3, 2018·Molecular Biology Reports·João Henrique Barbosa ToscanoAna Carolina de Souza Chagas
Sep 16, 2020·Animals : an Open Access Journal From MDPI·Linjie WangYan Wang

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Datasets Mentioned

BETA
DQ386890

Methods Mentioned

BETA
electrophoresis
PCR

Software Mentioned

Bestkeeper
SAS
geNorm
primer5
BLAST
NormFinder

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