title: Evaluation of housekeeping genes for normalizing real-time quantitative PCR assays in pig skeletal muscle at multiple developmental stages.
abstract: Quantitative PCR (qPCR) requires a constantly expressed housekeeping gene as an internal control, the expression of which is similar in different biological samples. In the present study, we evaluated the applicability and compared the consistency of the gene expression of 16 reference genes, i.e., 10 common and 6 candidate genes, through qPCR assays in pig skeletal muscles at multiple developmental stages. The stability of these 16 potential reference genes was investigated using the geNorm and NormFinder methods. Our results indicated that DRAP1 and RNF7 were the most appropriate combination to normalize gene expression in the Yorkshire samples, the combination of DRAP1 and WSB2 were appropriate in the Tongcheng samples, H3F3A and DRAP1 in prenatal periods, DRAP1 and RNF7 in postnatal periods, and the combination of DRAP1 and WSB2 was most suitable for accurate normalization in whole samples. This result provides a strong basis for similar studies in pigs that explore the molecular mechanisms underlying skeletal muscle development.
authors: Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong (Central China) Agricultural University, Wuhan, Hubei, PR China.
authors: Yueying
authors: Y
authors: Wang
authors: Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong (Central China) Agricultural University, Wuhan, Hubei, PR China.
authors: Yuqiang
authors: Y
authors: Zhao
authors: Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong (Central China) Agricultural University, Wuhan, Hubei, PR China.
authors: Ji
authors: J
authors: Li
authors: Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong (Central China) Agricultural University, Wuhan, Hubei, PR China.
authors: Huijing
authors: H
authors: Liu
authors: Department of Animal Science, Michigan State University, East Lansing, MI, United States.
authors: Catherine W
authors: CW
authors: Ernst
authors: China Education Ministry's Key Laboratory in Agricultural Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, PR China.
authors: Xiaoran
authors: X
authors: Liu
authors: Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong (Central China) Agricultural University, Wuhan, Hubei, PR China.
authors: Guorong
authors: G
authors: Liu
authors: Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong (Central China) Agricultural University, Wuhan, Hubei, PR China.
authors: Yu
authors: Y
authors: Xi
authors: Key Lab of Swine Genetics and Breeding of Ministry of Agriculture, Key Lab of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong (Central China) Agricultural University, Wuhan, Hubei, PR China. Electronic address: leimg@mail.hzau.edu.cn.
authors: Minggang
authors: M
authors: Lei
mesh_terms: D000818:Animals; D020869:Gene Expression Profiling / Q000379:methods; D020043:Genes, Essential / Q000235:genetics*; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics; D018482:Muscle, Skeletal / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D012097:Repressor Proteins / Q000235:genetics; D013552:Swine / Q000235:genetics*; D044767:Ubiquitin-Protein Ligases / Q000235:genetics
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D012097:Repressor Proteins; D044767:Ubiquitin-Protein Ligases
keywords: Multiple developmental stages; Pig; Reference gene; Skeletal muscle; q-PCR