分析与解决PCR扩增肠癌组织P53基因时失败的问题,为如何解决PCR技术在科研应用中的问题提供一个实例.方法 经初步分析,将失败原因确定为DNA模板有问题、采用下列方法处理模板再进行PCR反应:用70%乙醇再清洗模板、用新标本重新提取模板、用PCR反应成功与失败的模板配成混合模板.比较PCR成功与失败所用模板的差异,寻找与证实失败的原因.结果 最终发现溶解DNA模板的TE缓冲液浓度过高,使PCR扩增失败,排除之后获得成功.结论 当PCR扩增失败时,不妨从最简单的反应体系的离子环境先找原因.“,” objective To analyze and resolve the problem failing to amplify p53 gene by PCR in colonic cancer and provide a example of resloving problems in application of PCR in scientific research. Methods After analysis the failure cause was believed to be of the DNA template. The following methods were used to process the template and then new PCR were performed washing the template again with 70% ethanol extracting DNA template again from new samples mixing the template succeeded in PCR and the template failed in PCR as a mew template. The differences between the succeeded template and failed template were compared for identifying the cause. Results Finally the cause frustrating PCR was found to be the concentration of TE buffer used to dissolve the template too high. After the error was corrected PCR succeeded. Conclusion When PCR frustrating verifying the ion condition in reaction system may be the first thing to do. Key word