| 实时荧光定量RT-PCR检测siRNA表达载体抑制人肾癌786-0细胞G250 mRNA的表达 |
| |
| 引用本文： | 赵俊峰,郑少斌,赵善超,姜耀东,肖耀军,杨旭凯.实时荧光定量RT-PCR检测siRNA表达载体抑制人肾癌786-0细胞G250 mRNA的表达[J].中国实验诊断学,2011,15(3):404-406. |
| |
| 作者姓名： | 赵俊峰 郑少斌 赵善超 姜耀东 肖耀军 杨旭凯 |
| |
| 作者单位： | 1. 河南省中医院、河南中医学院第二附属医院,泌尿外科,河南,郑州,450002
2. 南方医科大学南方医院,泌尿外科,广东,广州,510515 |
| |
| 摘 要： | 目的评价实时荧光定量逆转录聚合酶链反应（RT-PCR）法检测小干扰RNA（siRNA）表达载体抑制人肾癌786-0细胞G250 mRNA表达的可靠性,以建立稳定的RNAi技术。方法体外合成四条针对G250基因的siRNA,并设计阴性对照和空白对照,转染肾癌786-0细胞株;根据G250 mRNA的序列设计特异性引物,荧光染料SYBR Green Ⅰ法定量RT-PCR测定G250 mRNA的表达情况。结果四条siRNA分别使G250 mRNA表达量降低了35.56%、49.27%、45.88%、65.13%,阴性对照和空白对照组无明显变化。结论实时荧光定量RT-PCR技术可以准确地检测mRNA的表达,可以用来评价siRNA的有效性。
|
| 关 键 词： | siRNA G250基因 肾细胞癌 实时荧光定量逆转录聚合酶链反应 |
Detecting of G250 mRNA in renal cell carcinoma 786-0 cell line by siRNA vector use real-time fluorescence quantitative RT-PCR assay |
| |
| ZHAO Jun-feng,ZHENG Shao-bin,ZHAO Shan-chao,et al..Detecting of G250 mRNA in renal cell carcinoma 786-0 cell line by siRNA vector use real-time fluorescence quantitative RT-PCR assay[J].Chinese Journal of Laboratory Diagnosis,2011,15(3):404-406. |
| |
| Authors: | ZHAO Jun-feng ZHENG Shao-bin ZHAO Shan-chao |
| |
| Institution: | ZHAO Jun-feng1,ZHENG Shao-bin2,ZHAO Shan-chao2,et al.(1.Department of Urology,Second affiliated Hospital of Henan Traditional Chinese Medicine University,Henan Zhengzhou 450002,China,2.Department of Urology,Nanfang Hospital,Southern Medical University,Guangzhou 510515,China) |
| |
| Abstract: | Objective To evaluate the reliability of real-time fluorescence quantitative RT-PCR in detecting G250 mRNA expression in G250 gene down-knocked renal cell carcinoma cell line 786-0 by RNA interference(RNAi) technique,in order to establish stable RNAi.Methods Four siRNAs against the G250 gene,negative control and blank were designed and transfected into 786-0 cells respectively,The specific primers were designed according to the sequence of G250 mRNA,and the fluorescence dye SYBR Green I were used for RT-PCR... |
| |
| Keywords: | siRNA |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ |
|
