GSM3996846: Sam_0062_lowantibiotic; Yersinia pseudotuberculosis IP 32953; Tn-Seq
Genomic DNA was extracted using ArchivePure DNA Extraction Kit, 5Prime Y. pseudotuberculosis strain IP32953 was previously used to generate a transposon library of approximately 40,000 unique insertion mutants using the Ez-Tn5 Kan2 transposome complex (Epicentre, UK) as described and validated in Willcocks et al (Willcocks SJ, Stabler RA, Atkins HS, Oyston PF, Wren BW. High-throughput analysis of Yersinia pseudotuberculosis gene essentiality in optimised in vitro conditions, and implications for the speciation of Yersinia pestis. BMC Microbiol. 2018;18(1):46.). Library preparation for Illumina MiSeq sequencing was performed as described in Willcocks et al (Willcocks SJ, Stabler RA, Atkins HS, Oyston PF, Wren BW. High-throughput analysis of Yersinia pseudotuberculosis gene essentiality in optimised in vitro conditions, and implications for the speciation of Yersinia pestis. BMC Microbiol. 2018;18(1):46.)