Utilization of a two-standard system in real-time PCR for quantification of gene expression in the brain.
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.
Naumenko
Vladimir S
VS
Institute of Cytology and Genetics, Siberian Division of the Russian Academy of Sciences, Novosibirsk, Russia. naumenko2002@mail.ru
Osipova
Daria V
DV
Kostina
Elena V
EV
Kulikov
Alexander V
AV
D000818:Animals; D001522:Behavior, Animal; D001923:Brain Chemistry / Q000235:genetics*; D002138:Calibration; D004247:DNA / Q000235:genetics*; D015870:Gene Expression / Q000502:physiology*; D008297:Male; D051379:Mice; D008806:Mice, Inbred AKR; D008808:Mice, Inbred CBA; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D044282:Receptor, Serotonin, 5-HT1A / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D014365:Tryptophan Hydroxylase / Q000235:genetics
D012333:RNA, Messenger; D044282:Receptor, Serotonin, 5-HT1A; D004247:DNA; C487802:Tph2 protein, mouse; D014365:Tryptophan Hydroxylase