加入自我延伸过程的融合PCR程序

The Fusion-PCR added Self-Extension Process

比较了加入自我延伸过程的融合PCR程序与传统PCR在扩增融合基因的扩增效果,自我延伸程序(94℃×1min,52℃×1min,72℃×1min)扩增2次,分别用不同的延伸时间:1min、2min、3min、5min,发现用2min、3min、5min延伸时间扩增出的融合基因条带比传统PCR显著亮一些,而用延伸时间为1min时,两种程序扩增出融合基因条带的亮度相近,说明自我延伸程序中的延伸时间是影响融合基因扩增量的关键因素。加入自我延伸过程的融合PCR扩增程序为:94℃×5min,(94℃×1min,52℃×1min,72℃×5min)×2次循环,(94℃×1min,52℃×1min,72℃×1min)×30次,4℃store。

In this paper, we discussed the fusion-PCR by adding self-extension process. The self-extension procedure was cycled for 2 times. When the extension length of self-extension procedure was 2, 3, or 5 minutes, the fusion-gene band amplified by fusion-PCR was significantly brighter than that of traditional PCR. However, when the extension length of the self-extension was 1 minutes, the genes amplified were similar between these two PCR processes. This indicates that the extension length of self-extension procedure is an im-portant factor influencing gene amplified in Fusion-PCR.