Human RORt mRNA time quantitative PCR method for detection and preliminary application.docVIP

 PAGE \* MERGEFORMAT 14 Human RORt mRNA time quantitative PCR method for detection and preliminary application OF: David Zweig Chen Jianguo Liu Yingzhao Wang Haisheng Wang Shengjun Ye Tian Shi Jie Yang prophets promise of the river with the best [Abstract] Objective: To establish a detection of human RORt (retinoid related orphan receptor gammat) mRNA in the SYBR Green real-time fluorescence quantitative PCR (real time fluorescence quantitative PCR, qRT PCR). Methods: According to the human RORt mRNA conserved sequence specific primers extracted by PMA and ionomycin stimulated human peripheral blood mononuclear cells (peripheral blood mononuclear cell, PBMC) total RNA, and reverse transcription into cDNA, and pMD 18T plasmid vector to construct the standard. by qRT PCR method for detection of RORt mRNA levels and to evaluate the method linearity, specificity and reproducibility. application of the method of detection of Graves disease (Graves’ disease, GD) patients in the RORt mRNA relative expression. Results: The standard curve correlation coefficient of 0.998, melting curve analysis showed a single peak, pMD18T RORt plasmid standard high and low copy number of the intra and inter-coefficients of variation were 6.5%, 8.4% and 9.6%, 11.2%. Preliminary studies indicate that GD patients was RORt mRNA relative expression levels were significantly higher than the control group (t =- 3.72, P lt;0.01). Conclusion: The detection of human RORt mRNA of SYBR Green real-time fluorescence quantitative PCR method is sensitive, specific and repeatable, provided the experimental basis for clinical application. [Keywords:] RORt; real-time quantitative PCR; SYBR Green [Abstract] Objective: To establish a SYBR Green real time fluorescent quantitative PCR (qRT PCR) method for the detection of human RORt (retinoid related orphan receptor gammat) mRNA.Methods: The specific primers were designed according to the conserved sequence of human RORt mRNA . Total RNA was