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Background  Real-time quantitative PCR (qPCR) is a method for rapid and reliable quantification of mRNA transcription. Internal standards such as reference genes are used to normalise mRNA levels between different samples for an exact comparison of mRNA transcription level. Selection of high quality reference genes is of crucial importance for the interpretation of data generated by real-time qPCR.    Results  In this study nine commonly used reference genes were investigated in 17 different pig tissues using real-time qPCR with SYBR green. The genes included beta-actin ( ACTB ), beta-2-microglobulin ( B2M ), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ), hydroxymethylbilane synthase ( HMBS ), hypoxanthine phosphoribosyltransferase 1 ( HPRT1 ), ribosomal protein L4 ( RPL4 ), succinate dehydrogenase complex subunit A ( SDHA ), TATA box binding protein ( TPB )and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein zeta polypeptide ( YWHAZ ). The stability of these reference genes in different pig tissues was investigated using the geNorm application. The range of expression stability in the genes analysed was (from the most stable to the least stable):  ACTB / RPL4 ,  TBP ,  HPRT ,  HMBS ,  YWHAZ ,  SDHA ,  B2M  and  GAPDH .    Conclusion  Expression stability varies greatly between genes.  ACTB, RPL4 ,  TPB  and  HPRT1  were found to have the highest stability across tissues. Based on both expression stability and expression level, our data suggest that  ACTB  and  RPL4  are good reference genes for high abundant transcripts while  TPB  and  HPRT1  are good reference genes for low abundant transcripts in expression studies across different pig tissues.

Selection of reference genes for gene expression studies in pig tissues using SYBR green qPCR