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不同参照基因在人海马组织表达研究中的比较。

A comparison between different reference genes for expression studies in human hippocampal tissue.

机构信息

Department of Medical Genetics, University of Campinas-UNICAMP, Campinas, SP, Brazil.

出版信息

J Neurosci Methods. 2012 Jun 30;208(1):44-7. doi: 10.1016/j.jneumeth.2012.04.020. Epub 2012 May 3.

Abstract

The reliability of gene expression studies by mRNA quantification is highly dependent upon several experimental procedures, including the choice of reference genes used for data normalization. In order to contribute to gene expression studies in mesial temporal lobe epilepsy (MTLE) we used microarray data, followed by real time quantitative PCR validation of selected housekeeping genes, to determine the most appropriate reference genes to be used in human hippocampal tissue gene expression studies. Our results unequivocally showed a significant impact of the reference gene chosen for normalization on the overall results of expression studies, clearly demonstrating the importance of adequate validation using stable reference genes. In addition, we found that HPRT, NSE, SDHA and SYP are suitable genes to be used as reference for normalization in expression studies of hippocampal tissue obtained from patients with MTLE.

摘要

mRNA 定量分析的基因表达研究的可靠性高度依赖于多个实验程序,包括用于数据归一化的参考基因的选择。为了有助于研究内侧颞叶癫痫 (MTLE) 中的基因表达,我们使用了微阵列数据,随后通过实时定量 PCR 对选定的管家基因进行验证,以确定最适合用于人类海马组织基因表达研究的参考基因。我们的结果明确表明,用于归一化的参考基因的选择对表达研究的整体结果有重大影响,清楚地表明了使用稳定的参考基因进行充分验证的重要性。此外,我们发现 HPRT、NSE、SDHA 和 SYP 是适用于作为 MTLE 患者海马组织表达研究中归一化的参考基因。

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