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目的为筛选食管癌相关的差异表达基因和基因组,建立哈萨克族食管癌表达文库。方法应用DAzLE系统构建哈萨克族食管癌cDNA表达文库,包括Trizol一步法提取总RNA,Oligo(dT)纤维素层析柱纯化mRNA,Super Script TMChoice System For cDNA Synthesis Kit合成cDNA,分级分离,纯化cDNA与质粒载体pSPORT1连接,电转化导入E.coliDH10B。结果14~18回收管合并,得到1.3×106个克隆,同时合成了肿瘤组和对照组的cDNA探针。结论成功构建的哈萨克族食管癌cDNA表达文库,可用于进一步筛选食管癌相关的差异表达基因,而且为搭建差异表达技术平台做好了准备工作。
Objective To screen esophageal cancer-related differentially expressed genes and genomes and establish a Kazakh esophageal cancer expression library. Methods The Kazak esophageal cancer cDNA expression library was constructed by DAzLE system. Total RNA was extracted by Trizol in one step, purified with Oligo (dT) cellulose column and cDNA was synthesized by Super Script TMChoice System For cDNA Synthesis Kit. The plasmid vector pSPORT1 was ligated and electrotransformed into E. coli DH10B. Results 14 to 18 tubes were pooled to obtain 1.3 × 106 clones. At the same time, cDNA probes of tumor group and control group were synthesized. Conclusion The successfully constructed Kazakh esophageal cancer cDNA expression library can be used to further screen esophageal cancer-related differentially expressed genes, and to prepare for the construction of differential expression technology platform.