title: Sample 1
description: Protocols: The Escherichia coli strain BW25113 large transposon mutant library described by Yasir et al. (2020) was used in this study. Briefly: Transposon Tnp001 is a mini-Tn5 transposon coding for kanamycin resistance (aph(3′)-Ia) that incorporates an outward oriented,IPTG-inducible tac promoter, 3′ to the kanamycin-resistance gene Genomic DNA was extracted using the Quick-DNA Fungal/Bacterial 96 Kit (Zymo Research). For LoRTIS sequencing library preparation,1 μg of genomic DNA was tagmented using an Illumina compatible MuSeek library preparation kit (Thermo Fisher Scientific) and the tagmented DNA was cleaned using 0.5X volume of AMPure XP magnetic beads (Beckman-Coulter). LongAmp Taq DNA polymerase (New England Biolabs, UK) was used with biotinylated oligonucleotide primers that hybridise specifically within the transposon to amplify DNA fragments containing transposon insertion junctions. A second primer that anneals to MuSeek adapters ('IonTMu-02') was used to generate biotinylated double-stranded PCR products containing transposon-genome junctions. These PCR products were enriched using a Dynabeads™ kilobaseBINDER™ kit (Thermo Fisher Scientific). A nested PCR reaction was carried out using NEB LongAmp Taq polymerase to generate DNA fragments with Oxford Nanopore multiplexing adapters that lacked biotin.
attributes.tag: ENA first public
attributes.value: 2022-05-30
attributes.tag: ENA last update
attributes.value: 2022-05-30
attributes.tag: External Id
attributes.value: SAMEA12548684
attributes.tag: INSDC center alias
attributes.value: Quadram Institute Bioscience
attributes.tag: INSDC center name
attributes.value: Quadram Institute Bioscience
attributes.tag: INSDC first public
attributes.value: 2022-05-30T12:16:20Z
attributes.tag: INSDC last update
attributes.value: 2022-05-30T12:16:20Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-11351:Sample 1
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: genotype
attributes.value: transformed with transposon Tnp001
attributes.tag: organism part
attributes.value: whole organism
attributes.tag: protocol
attributes.value: long reads library
attributes.tag: sample name
attributes.value: E-MTAB-11351:Sample 1