大豆DDRT分析中单引物造成假阳性的研究

摘要

[目的]探讨mRNA差异显示技术(DDRT)出现假阳性的原因.[方法]以大豆品种"吉林30"和"通农13"为材料,对DDRT分析中造成的假阳性进行了分析.[结果]DDRT假阳性的一个重要来源是由于单引物与cDNA组结合导致非特异性扩增造成的.在DDRT试验中,应平行设置单一引物的PCR试验以校正所得差异片段是否为假阳性或混杂有单引物PCR产物.[结论]为提高DDRT试验成功率奠定了基础.%[ Objective] The aim was to explore the reasons of false positives in Different Display Reverse Transcription (DDRT) analysis.[ Method] Soybean varieties “Jilin 30” and “Tongnong 13” were used as materials to carry out analysis on false positives in DDRT analysis.[ Result] An important origin of false positives appeared in DDRT analysis was the non-specific amplification caused by the combination of single primer and cDNA. The parallel PCR test of single primer should be set so as to verify whether the obtained fragments were the false positives or the PCR productions combined with single primer. [ Conclusion] This study had provided basis for improving the success rate of DDRT experiment.