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北京美亿美生物_科研技术服务平台 | mymbio.com Reviews
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北京美亿美生物_科研技术服务平台
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DGGE(denaturing gradient gel electrophoresis)，即变性梯度凝胶电泳，是根据DNA在不同浓度的变性剂中解链行为的不同而导致电泳迁移率发生变化，从而将片段大小相同而碱基组成不同的DNA片段分开。 引物设计，扩增，基因组DNA电泳图， PCR电泳图， DGGE电泳图，聚类分析图，主成份分析图和多样性分析 DGGE胶图主要条带标注，DGGE胶中特异条带的回收，特异条带PCR扩增，特异条带克隆测序，序列比对分析，种属系统发育分析.
北京美亿美生物_科研技术服务平台
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浏览次数 957次 更新时间 2013-04-15 22:51:10. 不同之处是Primer上要加一个GC夹 GC Clamp)，GC夹的序列为 CGCCCGCCGCGCGCGGCGGGCGGGGCGGGGGC. 1)新鲜配制的Ammonium Persulfate Solution APS 0.1g in 1ml H2O. 顶层胶 4ml 0%变性剂 4微升TEMED 40微升APS溶液. 底层胶 1ml 80%变性剂 4微升TEMED 40微升APS溶液. 40%变性剂凝胶溶液 6.5ml 0%变性剂 6.5ml 80%变性剂 13微升TEMED 130微升APS溶液. 60%变性剂凝胶溶液 3.25ml 0%变性剂 9.75ml 80%变性剂 13微升TEMED 130微升APS溶液. 用万分之一的EB或SYBR Green I染色, 我比较了一下发现EB效果明显优于SYBR Green I。
北京美亿美生物_科研技术服务平台
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浏览次数 887次 更新时间 2013-04-15 22:50:06. DGGE，CGGE及TGGE三种方法是基于相同的原理而设计的突变检测方法，它仍均是根据DNA的解链特性而设计.对于同一定序列组成或的DNA片段来说，它具有恒定的解链温度(Tm)，但若其序列发生改变时，Tm值亦发生改变，在含有变性因素(变性剂， 高温)的凝胶半进行电泳，当其比链解开形成分叉时，电泳迁移的速度就会改变.Tm值 全要取决于DNA的碱基组成，序列中出现单碱基替换时，Tm亦发生改变，电泳迁移率 亦改变，此即DGGE，CDGE及TGGE鉴定突变的技术基础. 1变性梯度凝胶电泳(denaturing gel gradient electrophoresis，DGGE). 2恒定变性胶电泳(Constant deratarared gel electrophoresis CDGE). 3温度梯度凝胶电泳(Temperature gradient gel electrophoresis TGGE).
北京美亿美生物_科研技术服务平台
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