| 加入自我延伸过程的融合PCR程序 |
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| 引用本文： | 刘亮伟. 加入自我延伸过程的融合PCR程序[J]. 勤云标准版测试, 2009, 29(18) |
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| 作者姓名： | 刘亮伟 |
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| 作者单位： | 刘亮伟,杨海玉,胡瑜(河南农业大学生命科学学院,郑州,450002);李相前(淮阴工学院,江苏淮安,223001)? |
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| 基金项目： | 河南省科技攻关项目"木聚糖酶热稳定性的全息酶工程定向改造",江苏省自然科学基金"海栖热袍菌极耐热葡聚糖内切酶分子定向进化"? |
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| 摘 要： | 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℃×1 min,72℃×1 min)扩增2次,分别用不同的延伸时间:1 min、2 min、3 min、5 min,发现用2 min、3 min、5 min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1 min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 ℃×5 min,(94℃×1 min,52℃×1 min,72℃×5 min)×2次循环,(94℃×1 min,52℃×1 min,72℃×1 min)×30次,4℃ store.
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| 关 键 词： | 自我延伸过程 融合PCR 延伸时间 融合基因 |
The Fusion-PCR added Self-Extension Process |
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| 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. |
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| Keywords: | Extensionzz Fusion-PCRzz lengthzz Genezz |
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