...Technology Pros Cons References Golden Gate Type II restriction enzymes Can assemble up to 10 modules in a single...2008 Gibson Cloning Gibson assembly method Single reaction, can assemble large DNA sequences, no restriction...same multiple cloning site (MCS). Modules are assembled one at a time by standard restriction enzyme digestion...upon ligation. As a result, once two modules are assembled next to each other, a third module cannot be placed...high-throughput cloning strategies available for assembling these modular types of constructs, and different...restriction enzyme-based). Well suited for the modular assembly of highly similar or repetitive sequences. Intermediate...called “cassettes”) can be readily reused in the assembly of any later construct (i.e.,functional cassettes...

...Gibson Assembly (Image from Plasmid 101: Gibson Assembly ) Back to Top Type IIS Assembly (Golden Gate & MoClo... next cloning project. Golden Gate Cloning (Image from Plasmid 101: Golden Gate Cloning ) Back to Top ...on TOPO cloning. Back to Top Gibson Assembly (Isothermal Assembly Reaction) Isothermal cloning, more commonly... MoClo) Type IIS systems, such as Golden Gate and Modular Cloning (MoClo) , take advantage of the unique... to assemble before you get started. Check out Addgene's website for easy to use MoClo and Golden Gate...Gibson assembly DNA fragments with 20-40 base pair homology at their ends can be easily ligated together... commonly known as Gibson assembly ( protocol ), takes advantage of the properties of 3 common molecular...

...Open Enzyme parts have C/E overhangs for Golden Gate assembly. Plasmids Explore the collection and order...Endy Lab Plasmids Jennifer Molloy Lab Plasmids Golden Gate cloning...

...al., 2007). Alternatively, if you prefer Golden Gate assembly, there are a series of plasmids from the...and Dinneny, J. R. (2013) ‘A robust family of Golden Gate Agrobacterium vectors for plant synthetic biology...Plasmids from the Tsuyoshi Nakagawa lab are binary gateway vectors and can be used for promoter swapping and...transformant with a single insertion because it will segregate 3:1 in the T2 generation. You also have no control...genotype in T1 than T2. Your transgene should not segregate in T1, but it probably will in T2. Laura Lee...PMC4426443. 4. Nakagawa, T. et al. (2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation...

...collection of reporter plasmids is amenable to Golden Gate assembly for turning the single-input reporters into... into multi-input cell classifiers in one assembly step. Find the miRNA reporter collection at Addgene...

... Disadvantages References Yamamoto Lab Golden Gate Assembly transfection One vector expresses Cas9...

...Strains Plasmids 101: TOPO Cloning Plasmids 101: Golden Gate Cloning Read all of our Plasmid Cloning Posts..., the more recently developed Gibson Assembly Cloning and Gateway® cloning technologies, as well as several...recombinant plasmid to propagate. Any cells that contained non-recombinant vectors (re-ligated empty vectors, ...ccdB aspect, Gateway takes advantage of the same principle that cells will not propagate while expressing...focus on a unique gene that makes the popular GatewayTM method possible: ccdB. But what is ccdB, what ...expressed the ccdB gene and were therefore unable to propagate because CcdA wasn’t available to counteract the... to thoroughly screen colonies for the insert. Gateway® technology (developed by InvitrogenTM) is essentially...

...to quickly assemble vectors and test different ideas. We are a huge fan of the Golden Gate based MoClo...expression method enables us to quickly screen and investigate transcription factor candidates and the regulation...

...update how we make and distribute kits, I helped assemble a crack team of Addgenies that includes: Michelle...particular function. Project team A Project Team is assembled to complete a series of tasks with precisely defined...a microscopist, a biostatistician, and that one golden-handed technician who can do a certain difficult...project indeed. Mission team A Mission Team is assembled to make progress towards open-ended goals whose...attacking this problem?”, try thinking “Who can I assemble to attack this problem from every angle?” Your...someone to coordinate, someone to decide, someone to delegate, etc. If you find that there is not someone (or...

...gRNAs are easily inserted using BbsI and Golden Gate Assembly. Either plasmid can be also linearized with...elements, like a repair template, using Gibson Assembly or restriction enzyme cloning. Streptomyces bacteria...Microbiome Bacteria with a Toolkit for Modular Assembly of Broad-Host-Range Plasmids. ACS Synth Biol 7...