NEBuilder®高保真 DNA 组装(NEB#E2621) 多种商业化的无缝克隆产品,如 NEBuilder®高保真 DNA 组装试剂、NEBGibson 组装®试剂,工作原理如下:PCR扩增目的基因;核酸外切酶消化插入片段和载体的 5’ 末端产生单链 3’ 突出端;连接酶、重组酶或体内修复作用形成磷酸二酯键来共价连接插入片段和载体。这些技术能在载...
将 Gibson 组装预混液产品用于电转化时,需要将反应液稀释 3 倍,并取 1 μl 用于转化。 DNA:如果 Gibson 组装反应液中所有 PCR 产物的总体积不超过 Gibson 组装反应体积的 20%,无需进行 PCR 产物纯化。PCR 产物体积过多会引入更多 PCR 产物中存在的 PCR 反应缓冲液和未使用的引物,从而降低 Gibson 组装和转...
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应用NEB®Golden Gate 进行复杂组装 Gibson 组装^{®}(NEB #E2611) Gibson 组装^{®}是在等温条件下,单管反应实现多个带重叠末端的 DNA 片段的组装,反应不会引入无关序列(无缝组装)。Gibson 组装^{®}预混液包含三种不同的酶,且在同一缓冲液中均有活性。将组装好的、完全连接的质粒转化到感受态细胞。...
Find out how NEBuilder® HiFi DNA Assembly can reliably join DNA fragments in a single tube, isothermal reaction, with advantages over NEB Gibson Assembly®. NEBUILDER® HIFI DNA ASSEMBLY® : Bridging dsDNA with a ssDNA Oligo Learn how NEBuilder® HiFi DNA Assembly bridges dsDNA ...
Find out how NEBuilder® HiFi DNA Assembly can reliably join DNA fragments in a single tube, isothermal reaction, with advantages over NEB Gibson Assembly®. Golden Gate Assembly Workflow Find out how Golden Gate Assembly can be used to quickly join multiple DNA fragments. What is a Type ...
3244 0 00:58 App Gibson 组装流程 117 0 01:48 App 限制性内切酶在液滴数字 PCR 中的应用 199 0 01:46 App 限制性内切酶在光学图谱中的应用 1159 0 06:42 App 从细胞&血液样本中提取高分子量 DNA 操作步骤 85 0 00:53 App 省时(TIME-SAVER™)限制性内切酶 ...
NEBuilder®高保真DNA组装基于Gibson无缝组装原理,并在其基础上进行了优化,可实现精准的DNA片段连接。试剂盒通过使用新型高保真聚合酶来实现简单快速的无缝克隆,是合成生物学的绝佳利器。 PART 01 选择NEBuilder®的4大理由: 1. 省时: 15分钟内完成简单快速的无缝克隆。
Daniel G. Gibson, of the J. Craig Venter Institute, described a robust exonuclease-based method to assemble DNA seamlessly and in the correct order, eponymously known as Gibson Assembly. The reaction is carried out under isothermal conditions using three enzymatic activities: a 5’ exonuclease gen...
Gibson Assembly has been successfully used to reliably join up to six DNA fragments into a single molecule. This remarkably straightforward and powerful techniques makes quick work of large multi-fragment assemblies but it is also useful for more routine applications such as cloning. The Gibson Assem...