摘要
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
出处
《江西医学检验》
2000年第3期132-134,共3页
Jiangxi Journal of Medical Laboratory Sciences
基金
江西省科委资助项目的部分内容!(课题编号 F940502)
