Objective To establish a real-time quantitative PCR (RT-qPCR) method to detect the transcription level of xanthine catalase/oxidase (XDH/XO) gene in Wistar rats, so as to quantitatively detect the XDH/XO gene at the transcription level.
Methods Total RNA was extracted from the liver tissue of Wistar rats, and cDNA was obtained by reverse transcription, which was diluted to 5 concentration gradients with a 10-fold dilution factor. The designed primer sequences and internal reference genes were used for RT-qPCR detection, and the expression of XDH/XO gene was obtained. the standard curve. Then, the changes of XDH/XO gene transcription level in Wistar rat hyperuricemia animal model were detected.
Results The standard curve of XDH/XO gene detected by RT-pPCR method showed a single dissolution peak, and R2 was close to 1, which could detect the changes in the transcription level of XDH/XO gene in hyperuric acid animal models.
Conclusion The RT-qPCR method for detecting the transcription level of XDH/XO gene in Wisar rats has the characteristics of quantitative accuracy and good repeatability, and can be applied to the pathogenesis of hyperuricemia and the research of new drugs.