Department of Toxicogenomics, National Institute of Public Health, Prague, Czech Republic.
Dis Markers. 2012;32(3):203-10. doi: 10.3233/DMA-2011-0875.
Histologically verified pairs (n=10) of pancreatic tumors and non-neoplastic tissues were used for quantitative real-time PCR and the stability of 24 reference genes was analyzed with geNorm and NormFinder software. Raw C{q} values correlated with the degree of RNA degradation. This correlation was abolished by normalization to C{q} of 18S endogenous control gene. Both geNorm and NormFinder programs suggested EIF2B1, ELF1, MRPL19, and POP4 as the same most stable genes. We have thus identified suitable reference genes for future expression studies in pancreatic carcinoma. Normalization method reducing the effects of RNA degradation on the quality of results was also developed.
采用组织学验证的配对（n=10）胰腺肿瘤和非肿瘤组织进行实时定量 PCR，并使用 geNorm 和 NormFinder 软件分析 24 个参考基因的稳定性。原始 Cq 值与 RNA 降解程度相关。通过标准化到内参基因 18S 的 Cq 值可消除这种相关性。geNorm 和 NormFinder 程序均建议 EIF2B1、ELF1、MRPL19 和 POP4 作为最稳定的基因。因此，我们已经确定了用于未来胰腺癌表达研究的合适参考基因。还开发了一种可以减少 RNA 降解对结果质量影响的归一化方法。
