Golden Gate assembly methods are also available for plants (Qi-Jun Chen and Yao-Guang Liu labs) and zebrafish (Wenbiao Chen lab). Multiple gRNA cassettes are assembled using BsaI-mediated Golden Gate cloning method.

Kit #1000000062: Production of … Golden Gate Assembly methods. Keywords: Pichia pastoris, Cell engineering, Golden Gate cloning, GoldenMOCS, Golden PiCS, Synthetic biology * Correspondence: brigitte.gasser@boku.ac.at 1Department of Biotechnology, BOKU University of Natural Resources and Life Sciences, Muthgasse 18, 1190 Vienna, Austria

article : doi: 10.1093/nar/gkt1361: ... CAG promoter-driven expression vector suitable for ligation of multiple inserts by Golden Gate Cloning using Esp3I. This page links to a number of protocol pages that will help you design DNA sequences for Golden Gate Assembly, learn about the various different Golden Gate Assembly techniques and when we employ each one, and conduct the different types of Golden Gate Assembly reactions. PubMed PMID 24954249. Golden Gate Assembly uses Type IIS restriction enzymes, which cleave outside of their … This cloning technique utilizes so-called type II restriction endonucleases.

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The inserts and cloning vectors are designed to place the Type IIS recognition site distal to the cleavage site, such that the Type IIS REase can remove the recognition sequence from the assembly. 1). The Golden GATEway cloning kit is a combinatorial approach that couples Golden Gate cloning with Multisite Gateway TM cloning. Using the type IIS restriction endonuclease BsaI and T4 DNA ligase, ready-to-use plant transformation

company name ... pCAG-Golden-Gate-Esp3I-Destination. It is based on the Golden Gate cloning method. This system was specifically designed for generating transgenesis constructs, but is also suitable for creating fusion proteins, and can be used in many different model organisms. Publication List for Kits: Kit #1000000055: Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. www.addgene.org help@addgene.org GreenGate Cloning Kit Description: GreenGate is a simple and efficient cloning system for rapidly assembling plant transformation constructs. help@addgene.org Golden GATEway Cloning Kit Description: The Golden GATEway cloning system combines Golden Gate and Multisite Gateway cloning for construction of complex plasmids in a predefined order.

Golden Gate Assembly This page serves as the main repository for everything Golden Gate Assembly-related. This “Golden Gate Modular Cloning (MoClo) Plant Parts Kit” can be found in Addgene along with the “Golden Gate MoClo Plant tool kit” that has all the vector backbones and sequences needed for domestication of new sequences and assembly into single and multigene binary constructs. home > Addgene > pCAG-Golden-Gate-Esp3I-Destination product summary. Golden Gate Assembly and its derivative methods exploit the ability of Type IIS restriction endonucleases (REases) to cleave DNA outside of the recognition sequence. Golden Gate assembly involves the simultaneous activities of a Type IIS restriction enzyme such as BsaI, and a DNA ligase such as T4 DNA ligase. This assembly is performed in vitro.Most commonly used Type IIS enzymes include BsaI, BsmBI, and BbsI. 86 talking about this. If you want to scale up to 3-7 gRNAs, Addgene has two Golden Gate Assembly-based mammalian systems from the Gersbach and Yamamoto labs.

GoldenPiCS, our Golden Gate-derived P. pastoris cloning system, consists of three hierarchical backbone (BB) levels for flexible generation of overexpression plasmids containing multiple transcription units (up to eight per plasmid), four different selection markers and five loci either for targeted genome integration or episomal plasmid maintenance (Fig. Golden Gate cloning was established in 2008 and has been widely used since (for example see 4–6). Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T. Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400.


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