Die Plant Science Student Conference (PSSC) wird seit 20 Jahren im jährlichen Wechsel von Studierenden der beiden Leibniz-Institute IPK und IPB organisiert. Im Interview erläutern Christina Wäsch (IPK) und Carolin Apel (IPB),…
Über 600 Gäste kamen am 4. Juli ans IPB zur Langen Nacht, die Wissen schafft, um bei unserem Wissenschafts-Quiz-Parcours viel Neues zu erfahren und ihre Kenntnisse unter Beweis zu stellen. Unser Programm in diesem Jahr…
Marillonnet, S.; Grützner, R.;Synthetic DNA Assembly Using Golden Gate Cloning and the Hierarchical Modular Cloning PipelineCurr. Protoc. Mol. Biol.130e115(2020)DOI: 10.1002/cpmb.115
Methods that enable the construction of recombinant DNA molecules are essential tools for biological research and biotechnology. Golden Gate cloning is used for assembly of multiple DNA fragments in a defined linear order in a recipient vector using a one‐pot assembly procedure. Golden Gate cloning is based on the use of a type IIS restriction enzyme for digestion of the DNA fragments and vector. Because restriction sites for the type IIS enzyme used for assembly must be present at the ends of the DNA fragments and vector but absent from all internal sequences, special care must be taken to prepare DNA fragments and the recipient vector with a structure suitable for assembly by Golden Gate cloning. In this article, protocols are presented for preparation of DNA fragments, modules, and vectors suitable for Golden Gate assembly cloning. Additional protocols are presented for assembly of defined parts in a transcription unit, as well as the stitching together of multiple transcription units into multigene constructs by the modular cloning (MoClo) pipeline.