动物实验,-荧光定量PCR z-bo 2020-08-21 587

　　OBJECTIVE: To compare the relative expression of xanthine dehydrogenase oxidase (XDH/XO) mRNA in macaque liver tissue, epithelial tissue, testis tissue, myocardial tissue, and kidney tissue, using fluorescence quantitative PCR and semi-quantitative PCR for comparison. Improve the methodology of experimental research to provide reference.

　　Method: Using the same primer sequence and internal reference gene, total RNA was extracted from fresh liver tissue, epithelial tissue, testis tissue, myocardial tissue and kidney tissue of healthy monkeys, and then subjected to fluorescence quantitative PCR and semi-quantitative determination respectively. Relative quantitative detection by PCR. Perform comparative analysis on the results.

　　Results: Both detection methods can detect the expression of XDH/XO mRNA in fresh liver tissue, epithelial tissue, testis tissue, myocardial tissue, and kidney tissue of healthy rhesus monkeys, and the sensitivity of fluorescence quantitative PCR is 0.4 in liver tissue and semi-quantitative PCR The sensitivity can detect the expression of XDH/XO mRNA in 15.625ng of total RNA in liver tissue. The sensitivity of fluorescent quantitative PCR is 39 times that of semi-quantitative PCR. The detection results of the two methods both showed that the expression of XDH/XO mRNA in different tissues of rhesus monkeys was highest in liver tissues. Fluorescence and semi-quantitative PCR produce different results in other tissues with low expression levels.

　　Conclusion: Both methods can be used to detect the relative expression of xanthine oxidase dehydrogenase (XDH/XO) mRNA in liver tissue, epithelial tissue, testis tissue, myocardial tissue and macaque tissue. The sensitivity of real-time quantitative PCR is higher than half Quantitative PCR method.