standardized-modular-assembly-polycistronic-operons-with-modular

Standardized Modular Assembly of Polycistronic Operons with Modular Cloning (MoClo) using the <em>In-Cloning</em> toolkit

Modular Cloning (MoClo) toolkits enable the rapid assembly of multigene constructs. They are based on Golden Gate cloning, which uses Type IIS restriction ...

Max Planck Institute for Terrestrial Microbiology

Synthetic Sc2.0 yeast strains contain hundreds to thousands of  recombination sites that allow restructuring of the  genome by SCRaMbLE. Thus, a highly diverse yeast population can arise from a single genotype. The selection of genetically diverse candidates with rearranged synthetic chromosomes for downstream analysis requires an efficient and straightforward workflow. Here we present loxTags, a set of qPCR primers for genotyping across  sites to detect not only deletions but also inversions and translocations after SCRaMbLE. To cope with the large number of amplicons, we generated qTagGer, a qPCR genotyping primer prediction tool. Using loxTag-based genotyping and long-read sequencing, we show that light-inducible Cre recombinase L-SCRaMbLE can efficiently generate diverse recombination events when applied to Sc2.0 strains containing a linear or a circular version of synthetic chromosome III.

Timon A. Lindeboom

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