age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues与年龄相关的变化相对常用的管家基因表达稳定选择猪组织.pdfVIP

Uddin et al. BMC Research Notes 2011, 4:441 /1756-0500/4/441 RESEARCH ARTICLE Open Access Age-related changes in relative expression stability of commonly used housekeeping genes in selected porcine tissues 1,2 1 1 1 1 Muhammad Jasim Uddin , Mehmet Ulas Cinar , Dawit Tesfaye , Christian Looft , Ernst Tholen and Karl Schellander1* Abstract Background: Gene expression analysis using real-time RT-PCR (qRT-PCR) is increasingly important in biological research due to the high-throughput and accuracy of qRT-PCR. For accurate and reliable gene expression analysis, normalization of gene expression data against housekeeping genes or internal control genes is required. The stability of reference genes has a tremendous effect on the results of relative quantification of gene expression by qRT-PCR. The expression stability of reference genes could vary according to tissues, age of individuals and experimental conditions. In the pig however, very little information is available on the expression stability of reference genes. The aim of this research was therefore to develop a new set of reference genes which can be used for normalization of mRNA expression data of genes expressed in varieties of porcine tissues at different ages. Results: The mRNA expression stability of nine commonly used reference genes (B2M, BLM, GAPDH, HPRT1, PPIA, RPL4, SDHA, TBP and YWHAZ) was determined in varieties of tissues collected from newborn, young and adult pigs. geNorm, NormFinder and BestKeeper software were used to rank the genes according to their stability. geNorm software revealed that RPL4, PPIA and YWHAZ showed high stability in newborn and adult pigs, while B2M, YWHAZ and SDHA showed high stability in you