PT  - JOURNAL ARTICLE
AU  - Hualan Liu
AU  - Morgan N. Price
AU  - Robert Jordan Waters
AU  - Jayashree Ray
AU  - Hans K. Carlson
AU  - Jacob S. Lamson
AU  - Romy Chakraborty
AU  - Adam P. Arkin
AU  - Adam M. Deutschbauer
TI  - Magic pools: parallel assessment of transposon delivery vectors in bacteria
AID  - 10.1101/158840
DP  - 2017 Jan 01
TA  - bioRxiv
PG  - 158840
4099  - http://biorxiv.org/content/early/2017/07/03/158840.short
4100  - http://biorxiv.org/content/early/2017/07/03/158840.full
AB  - Transposon mutagenesis coupled to next-generation sequencing (TnSeq) is a powerful approach for discovering the functions of bacterial genes. However, the development of a suitable TnSeq strategy for a given bacterium can be costly and time-consuming. To meet this challenge, we describe a parts-based strategy for constructing libraries of hundreds of transposon delivery vectors, which we term “magic pools”. Within a magic pool, each transposon vector has a different combination of promoters and antibiotic resistance markers as well as a random DNA barcode sequence, which allows the tracking of each vector during mutagenesis experiments. To identify an efficient vector for a given bacterium, we mutagenize it with a magic pool and sequence the resulting insertions; we then use the best vector to generate a large mutant library. We used the magic pool strategy to construct transposon mutant libraries in five genera of bacteria, including three genera of the phylum Bacteroidetes.