Utilization of a two-standard system in real-time PCR for quantification of gene expression in the brain

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• 作者：Vladimir S. Naumenko ; Daria V. Osipova ; Elena V. Kostina ; Alex ; er V. Kulikov
• 关键词：5-HT_1A receptor ; Tph2 ; mRNA level ; RT-PCR ; Real-time RT-PCR ; Catalepsy
• 刊名：Journal of Neuroscience Methods
• 出版年：2008
• 期刊代码：33_01650270
• 类别：neu
• 出版时间：30 May 2008
• 卷：170
• 期：2
• 页码：197-203
• 文件大小：201 K

摘要

In this study, we applied for real-time PCR the two-standard system that we had worked out previously for PCR with gel-detection of products. Genomic DNA of a known concentration was used as external standard and mRNA of the DNA-dependent RNA-polymerase II was used as internal standard. It was shown that PCR with gel-detection of products and real-time PCR provide similar results and demonstrate almost identical accuracy and repeatability when the two-standard system is used. With the help of the both methods and using the two-standard system we have confirmed the link between the genetically determined freezing reaction in mice and reduced 5-HT_1A receptor mRNA level in the midbrain. We have also found that the genetically determined freezing reaction in mice is not connected with changes in Tph2 gene expression.