| To normalize a set of quantitative real-time PCR (q-PCR) data, it is essential to determine an optimal number/set of housekeeping genes, as the abundance of housekeeping genes can vary across tissues or cells during different developmental stages, or ever under certain environmental conditions. In this study, out of 20 commonly used endogenous control genes,13,18, and 17 genes exhibited credible stability in 56 different tissues, ten types of adipose tissue, and five types of muscle tissue, respectively.Our analysis clearly showed that HSPCB/TOP2B is the most stable gene pair (M=0.67),however HMBS (M=2.05) stability is the last one in all 56 differernt tissues.HSPCB/TOP2B exhibited best stability (M=0.41) but HMBS(M=1.53) is the least stable gene in ten types of adipose tissues, in five types of muscle tissues HSPCB/TOP2B exhibited best stability (M=0.32),and 5S rRNA performed as the least stable gene.HSPCB/TOP2B are tow most stable genes both in 56 different tissues and five types of muscle tissues.During the optimal gene number analysis, our result showed that the optimal gene number is 16 (V=0.11) in 56 different tissues,and 11 genes exhibited the least V value (v=0.07) in 10 types of adipose tissues,15 genes are the best number in five types of muscle tissues.To verify if it is enough to normalize a set of q-PCR data using three housekeeping genes.we analysised the relation between NF3 and NFopt. It shows that the is highly minus related between them, which r>0.94. In 56 different tissues r=0.94, r=0.98 in adipose tissues,and muscle types tissues r=0.99.So these support our conclusions.In summary, we advice in 56 different tissues,TOP2B, HSPCB and YWHAZ can use to normalize q-PCR data,and in adipose type tissues are HSPCB,ALDOA,GAPDH,while TOP2B,HSPCB,YWHAZ are suitable for muscle tissues. Our report will serve as a valuable reference for other studies aimed at measuring tissue-specific mRNA abundance in porcine samples. |
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