地高辛随机引物法标记探针的Southern杂交技术优化

Technological Improvement of Southern Blot Using Digoxigenin-labeled Probes with Random-primed Method

为检测转基因抗虫棉中的Bt Cry1A基因的表达,采用地高辛随机引物法标记探针、化学发光检测的Southern杂交技术,通过纯化探针、使用最适探针浓度、优化真空转印、杂交及免疫检测方法等一系列优化过程,最终得到背景低、信号强的Southern杂交结果。

Southern blot using digoxigenin（ DIG ） -labeled probes with random-primed method and chemiluminescent detection was used to detect Bt CrylA gene of transgenic insect-resistant cotton. A serial of procedure was modified by purifying the DIG-labeled probes, using optimum probes concentration, improving the process of vacuum transferring, hybridization and immunoassay. Finally, we achieved good Southern blot result with low background and high sensitivity.