期刊
JOURNAL OF BIOTECHNOLOGY
卷 150, 期 3, 页码 288-293出版社
ELSEVIER
DOI: 10.1016/j.jbiotec.2010.09.949
关键词
Skeletal muscle; Reference genes; qPCR; geNorm; Normfinder; ROCK2
资金
- National Key Foundation Research and Development Program of China [2006CB102102]
- National High Technology Research and Development Program [2006AA10Z1D6]
- Key and Specific National Project for Creating New Biological Species Transgenically [2008ZX08006-002]
- National Natural Science Foundation of China [30800782]
- New Teacher Research Fund for the Doctoral Program of Higher Education [20070504064]
Quantitative PCR (qPCR) is a method for rapid and reliable quantification of mRNA. Internal controls such as reference genes are used to normalize mRNA levels between different samples for an exact comparison of gene transcription level. However, the expression levels of these reference genes may vary between cell types, developmental stages, species and experimental conditions, thus proper normalization strategy is an important precondition for reliable conclusions. In this study, we explored 10 commonly used reference genes in porcine skeletal muscle using SYBR green qPCR. We used both geNorm and NormFinder to analyze the expression stability and found that PPIA, HPRT and eEF-1 gamma were suitable internal controls for porcine skeletal muscle. However, PPIA, HPRT and SDHA were suitable for skeletal muscle of western pigs while PPIA, eEF-1 gamma and HPRT for indigenous Chinese pigs. Normalized qPCR data of ROCK2 were compared with microarray data to evaluate our developed set of reference genes. (C) 2010 Elsevier B.V. All rights reserved.
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