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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (18): 96-98.

• 食品 营养 检测 安全 • 上一篇    下一篇

加入自我延伸过程的融合PCR程序

刘亮伟1,杨海玉1,胡 瑜1,李相前2   

  • 收稿日期:2009-04-10 修回日期:2009-05-06 出版日期:2009-09-20 发布日期:2009-09-20
  • 基金资助:

    河南省科技攻关项目;江苏省自然科学基金

The Fusion-PCR added Self-Extension Process

刘亮伟1，杨海玉1，胡 瑜1，李相前2   

  • Received:2009-04-10 Revised:2009-05-06 Online:2009-09-20 Published:2009-09-20

PDF (PC)

1898

摘要：

本文探讨了加入自我延伸过程的融合PCR程序与传统PCR扩增条带效果，自我延伸程序（94*1，52*1，72*）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。

关键词: 玉米, 玉米, 同工酶, 自交系

Abstract:

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 important factor influencing gene amplified in Fusion-PCR.