高纯化RNA分离方法的建立与应用  

Method for Separating RNA of High Purity

作　　者：饶国洲[1] 李昂[1] 景娟[1] 姚天华[1] 石建锋[1] 

机构地区：[1]西安交通大学口腔医学院中心实验室,陕西西安710004

出　　处：《现代检验医学杂志》2005年第6期4-6,共3页Journal of Modern Laboratory Medicine

摘　　要：目的建立一种从不同生物体来源的组织材料中进行简单、快速、高纯化分离RNA的方法.方法利用胍盐裂解并抑制RNA的降解,用特异的RNA吸附剂吸附;通过清洗液的清洗和洗脱液洗脱,获得高纯化RNA.并用该方法在HL-60细胞中分离总RNA,进行RT-PCR扩增Human TFR(Transferrin Receptor)基因.结果RNA总得率大于80%;纯度高(A260/2801.9～2.0);产量稳定(平均10～15 μg/1×106细胞,1～5μg/mg组织,2.5～5μg/ml全血,50～100 μg/1×109细菌,10～20μg/1×108酵母菌);时间短(20～45 min),无基因组DNA污染,RNA降解少,完整性较好.Human TFR RT-PCR扩增电泳结果显示出1.0kbp,2.0kbp,4.4kbp的DNA片段.结论该方法操作简便、快速,适应于普通实验室对RNA的分析研究工作.Objective The experiment was conducted in order to found a method to separate RNA of high purity from tissues of different organisms simply and rapidly. Methods Schizolysised and suppressed the degradation of RNA with guanidine salt and absorbed the production with the specific absorbent,then RNA of high purity could be got after washing and dialysing the production with the detergent and dialysate. In this experiment,separated the total RNA from HL-60 cell strain through the method mentioned above,and amplified Human TFR （Transferrin Receptor）gene by RT-PCR. Results The total RNA obtained was of high purity and its percentage was more than 80%, （A260/280 1. 9～2. 0）. The average amount of them which was got from different organisms were as follow：10～15μg/1 × 10^6 Cells, 1～5μg/mg Tissue, 2. 5～5μg/ml Blood, 50～100μg/1× 10^9 Bacteria and 10～ 20μg/1 × 10^8 Yeast. The production was of high purity,low degradation and not mixed with genomic DNA. It took only 20～45 min to finish the whole experiment. Finally got TFR gene of 1. 0 kbp,2.0 kbp and 4. 4 kbp in TR-PCR amplified electrophoresis. Conclusion This method is simple,rapid and suited for general laboratory studies of RNA analysis.

关 键 词：RNA 纯化分离 方法 

分 类 号：Q503[生物学—生物化学]