逆转录法制备地高辛标记的食管癌cDNA探针

Preparation of esophageal carcinoma cDNA probe labeled with digoxigenin by reverse transcription reaction

目的 :制备食管癌cDNA探针用于ReverseNorthernBlot筛选差异基因片段。方法 :提取食管癌组织总RNA ,通过逆转录反应 ,用地高辛标记为cDNA探针。结果 :所制备的探针灵敏度高达 0 .0 1ng/L。结论 :用该方法制备cDNA探针快速、经济 ,所需模板RNA量少 ,可用于大规模初筛差异基因片段的研究。

Aim: To prepare the esophageal carcinoma cDNA probe for reverse northern blot in screening the differential gene fragments.Method: The cDNA probe of esophageal cancer tissue was labeled with digoxigenin by reverse transcription reaction, and applied in reverse northern blot to initially screen the retrieved differential cDNA fragments. Results: Sensitivity of the probe was as high as 0.01 ng/L. Conclusions: It is a rapid economical technique to prepare cDNA probe, and might be used in large scale screening of the differential fragments.