Reference: Ip K, et al. (2020) High-Throughput DNA Assembly Using Yeast Homologous Recombination. Methods Mol Biol 2205:79-89

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Abstract


Yeast homologous recombination is a reliable, low-cost, and efficient method for DNA assembly. Using homology regions as short as 24 base pairs, constructs of up to 12 unique parts can be assembled into a diverse range of vectors. The simplicity and robustness of this protocol make it amenable to laboratory automation and high-throughput operations. Here we describe a high-throughput protocol to generate DNA parts through PCR, assemble them into a vector via yeast transformation, and "shuttle" the resulting plasmid constructs into E. coli for storage and propagation. Though this protocol is intended for high-throughput workflows, it can be easily adapted for bench-scale DNA assembly.

Reference Type
Journal Article
Authors
Ip K, Yadin R, George KW
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