|
This test consists of three parts, the stability of the intestinal tissue reference gene the different lactation of mouse mammary tissue, liver tissue of mice and young mice after different immune growing up. By fluorescence real-time quantitative PCR gene B2M, ACTB, GAPDH, SDHA, HPRT1 and ARBP expression levels and applications geNorm program analysis, the final selection of a suitable reference gene, to lay the foundation for the study the expression of the target gene. Test: real-time quantitative PCR (qPCR) to detect one of the most commonly used technique in the amount of mRNA expression in the cells and tissues, and qPCR requires standardized data. By qPCR studies in the test six potential reference genes (B2M, ACTB, GAPDH, SDHA, HPRT1 and ARBP) expression in the mouse mammary tissues of different lactation. After SAS6.12 ANOVA model for statistical analysis, the results showed that the the B2M significant difference (P lt; 0.05). After geNorm program analysis, the selected reference gene stability high to low Sort GAPDH/HPRT1, ARBP, ACTB, SDHA, B2M. Thus recommended the application of gene GAPDH and HPRT1 as reference within the real-time quantitative PCR lactation mouse mammary tissue. Test: real-time quantitative PCR technology has been widely used in the detection and quantification of the cell or tissue mRNA transcription level. Select the appropriate reference gene can eliminate the different specimens there may be differences in RNA yield, quality and reverse transcription efficiency, so as to obtain the target gene-specific expression of the real differences. The application of this test real-time quantitative PCR study in mice after immune stimulation, B2M, ACTB, GAPDH, SDHA, HPRT1 and ARBP total of six reference gene expression in the liver tissue. The results show that the, six reference gene expression differences. The geNorm program statistical analysis to determine the ACTB, GAPDH housekeeping gene applies to the amount of the correction target gene expression, and lay the foundation for the study of mice immune stimulation liver target gene expression. Test: reference gene is commonly used in real-time quantitative PCR detection of mRNA expression levels of correction and standardization, but the reference gene expression by physiological stage, tissue or cell, as well as the experimental conditions. Therefore, this test select B2M, ACTB, GAPDH, SDHA, HPRT1 and ARBP a total of six reference gene, its expression in the young mouse small intestine tissue. After analysis geNorm program and NormFinder of procedures to finalize the reference gene SDHA and HPRT1 suitable for the the correction target gene expression, and to lay the foundation for the study of young mice intestinal tissue gene expression.
|