分子克隆之Golden Gate cloning
Golden Gate 克隆技术依赖于 1996 年首次发现的 IIS 型限制性内切酶。 IIS 型限制性内切酶与“传统”限制性内切酶的独特之处在于它们在识别序列之外切割,产生四个碱基侧翼突出端。由于这些突出端不是识别序列的一部分,因此可以定制它们以指导 DNA 片段的组装。如果设计正确,识别位点不会出现在最终构建体中,从而实现精...
推荐你几篇文献：都是做这个方向的Assembly of designer TAL effectors by Golden Gate cloning和Efficient design and assembly of custom TALEN and other TAL effector-based constructs for DNA targeting还有Generation of families of construct variants using golden gate shuffling和 A one pot, one ...
Golden Gate克隆示意图 下图描述了两种常用的插入片段制备办法,无论是酶切还是PCR,其根本还是要产生互补的粘性末端,因此只要粘性末端设计得当,可同时插入多种片段。具体的方法可参考2020年新发表的文章:Synthetic DNA Assembly Using Golden Gate Cloning and the Hierarchical Modular Cloning Pipeline,具体信息在文末给...
Learn the principles of Golden Gate cloning and how to simulate standard or batch Golden Gate cloning.
Golden Gate Shuffling: A One-Pot DNA Shuffling Method Based on Type IIs Restriction Enzymes Assembly of Designer TAL Effectors by Golden Gate Cloning Multiple-support response spectrum analysis of the golden gate bridge MULTIPLE-SUPPORT RESPONSE SPECTRUM ANALYSIS OF THE GOLDEN GATE BRIDGE. ...
Golden Gate cloning provides a precision module-based cloning technique for facile assembly of multiple genes in one construct. We present here a versatile resource for plant biologists comprising a set of cloning vectors and 96 standardized parts to enable Golden Gate construction of multigene ...
Golden Gate MoClo standards cloning restriction enzyme Introduction Custom DNA constructs play a fundamental role in biological research as cheap, easy-to-manipulate carriers of genetic information.1 Their applications include targeted genome editing,2 the expression of recombinant proteins,3 the construction...
Golden Gate Assembly Protocol for using NEBridge® Golden Gate Assembly Kit (BsmBI-v2) (NEB #E1602) 3.现在RNAi所用的shRNA则需要用Gibson Assembly Gibson Assembly® Gibson Assembly Cloning - addgene Gibson Assembly® Master Mix – Assembly (E2611) ...
Cite this protocol Maciá Valero, A., Prins, R.C., de Vroet, T., Billerbeck, S. (2025). Correction to: Combining Oligo Pools and Golden Gate Cloning to Create Protein Variant Libraries or Guide RNA Libraries for CRISPR Applications. In: Schindler, D. (eds) Golden Gate Cloning. Method...