长期冻存和新鲜胃癌组织总RNA提取方法的比较-中国组织工程研究.PDF

中国组织工程研究 第 16 卷 第24 期 2012 –06 –10 出版 Chinese Journal of Tissue Engineering Research June 10, 2012 Vol.16, No.24 长期冻存和新鲜胃癌组织总RNA提取方法的比较*★ 叶文静，韩纪举，赵晓民，夏作理 Comparison of total RNA extraction methods from long-term frozen and fresh gastric carcinoma tissues Ye Wen-jing, Han Ji-ju, Zhao Xiao-min, Xia Zuo-li Abstract Taishan Medical BACKGROUND: At present, the most reliable and commonly used methods are guanidine isothiocyanate based RNA College, Taian isolation method (typically Trizol) and the silica-gel column based RNA isolation method. In case of clinical rare tissues, it 271000, Shandong Province, China is necessary to evaluate the reliability and repetitiveness of the optional RNA isolation methods. Ye Wen-jing ★, OBJECTIVE: To compare the silica-gel column based RNA extraction method and Trizol method to determine which Master, Lecturer, technique is preferable when frozen, long-term stored or fresh gastric caricinoma tissues have to be evaluated for the Taishan Medical downstream molecular analysis. College, Taian 271000, Shandong METHODS: Frozen gastric carcinoma tissues were prepared by long-term storage (more than 2 years) in -80 ℃ while Province, China yewj75@126.com fresh tissues were harvested and processed immediately. The purity of RNA was determined with a spectrop hotometer whereas the Ct values of target sequences were recorded by quantitative reverse transcription-PCR (qRT-PCR) system to Supported by: the indirectly assess the RNA intactness (28S and 18S) and mRNA fragment sizes. Youth Foundation of Taishan Medical RESULTS AND CONCLUSION: The purity of RNA extracted with silica-gel column based and Trizol method was College* comparable. qRT-PCR data revealed that lower mean Ct values of 28 S