Hi Jonas,

I am a huge fan of the Modular Cloning Strategy. If you get the two plate kits (MoClo Toolkit, MoClo PlantParts) from Addgene, you will have a very solid synthetic biology starter kit in the lab. In my case, I have discovered 2-3 cross-contaminated modules (and other people say the same), but it's nonetheless amazing. The assembly works beautifully, very few wrong clones, the generation of new modules does also work quite well (considering that your gene does not have excessive numbers of BsaI or BpiI restriction sites), and possibilities and flexibility are just about unlimited. I must admit that I do not have experience with the other two. But GoldenBraid is iterative for multigene constructs, so more time-consuming. And the GreenGate kit also does not provide such an ample collection of readily available modules such as MoClo, and as far as I can see, the intermediates for multigene constructs are not plant transformation vectors, so these cannot be tested in planta on their own. Most importantly, the MoClo system is gaining more and more momentum, so that exchange of modules between different groups will also be a valauble ressource in the future.

As you say, in the end it depends on your specific needs. But the MoClo is great, no reservations, highly recommended!

Best wishes,

Johannes

 Dear Johannes,

Thank you very much for your extensive answer! The popularity of the cloning method is indeed an important consideration to take into account, I will have a talk with my promoter so we can order a kit (or preferably both kits) as soon as possible.

Thanks again!

Kind regards, Jonas