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

被引：88

作者：

Naumenko, Vladimir S. ^{[1
]}

Osipova, Daria V. ^{[1
]}

Kostina, Elena V. ^{[2
]}

Kulikov, Alexander V. ^{[1
]}

机构：

[1] Russian Acad Sci, Inst Cytol & Genet, Siberian Div, Lab Behav Neurogen, Novosibirsk 630090, Russia

[2] Russian Acad Sci, Siberian Div, Inst Chem Biol & Fundamental Med, Novosibirsk 630090, Russia

来源：

JOURNAL OF NEUROSCIENCE METHODS | 2008年 / 170卷 / 02期

关键词：

5-HT1A receptor; Tph2; mRNA level; RT-PCR; real-time RT-PCR; catalepsy;

D O I：

10.1016/j.jneumeth.2008.01.008

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

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-HT1A 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. (C) 2008 Elsevier B.V. All rights reserved.

引用

页码：197 / 203

页数：7

共 28 条

[1] Quantitation of hepatitis B virus genomic DNA by real-time detection PCR [J].