摘要
目的比较不同方法下的聚合酶链反应对PCR扩增的影响。方法对124例晚期乳腺癌外周血标本的CyclinD1基因进行PCR扩增。在加入DNA模板前分别用离心法和微型旋涡混合器振荡法混合PCR反应液,分装各管后加DNA模板进行扩增、电泳。结果用微型离心机离心PCR反应液后仍扩增不出或极少能扩出PCR产物,其阳性率为11.3%,且易污染;而用微型旋涡混合器振荡法混合PCR反应液后,PCR产物扩增良好(阳性率达99.2%),且不易污染。结论微型旋涡振荡法混合PCR反应液法能在减少污染的同时大大提高PCR阳性率,是一种切实可行的基因扩增方法。
Objective To investigate the influence of different mix method on PCR reaction mixture solution. Methods CyclinD1 gene was amplified by PCR with peripheral blood samples in 124 cases of advanced breast cancer. PCR reaction mixture solution was mixed by minicentrifuge DNA temple added. DNA temple was added in each tube and and minihelicoid agitation mixer respectively before electrophoresis was taken. Results The method of minicentrifuge mixed PCR reaction mixture solution could not or little amplify the PCR production. The positive rate was 11.3% and easily contaminated. Whereas with the method of minihelicoid agitation mixed PCR reaction mixture solution showed a good PCR production amplification and the band was clear, not easy contaminated and the positive rate was reached to 99.2%. Conclusion Because of the different mechanisms, the different mixer showsdifferent PCR positive rates. The minihelicoid agitation mixer mixed PCR reaction solution showed a good PCR production amplification rate and less contamination. It is a worthy method for gene PCR amplification.
出处
《青海医学院学报》
CAS
2007年第1期12-14,共3页
Journal of Qinghai Medical College
