characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-05-30T12:16:20Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-05-30T12:16:20Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA12548685
characteristics.characteristic: External Id
characteristics.text: Quadram Institute Bioscience
characteristics.characteristic: INSDC center name
characteristics.text: 2022-05-30T12:16:20Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-05-30T12:16:20Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS10155235
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-11351:Sample 2
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.characteristic: broker name
characteristics.text: 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.
characteristics.characteristic: description
characteristics.text: transformed with transposon Tnp001
characteristics.characteristic: genotype
characteristics.text: Escherichia coli BW25113
characteristics.characteristic: organism
characteristics.text: Escherichia coli BW25113
characteristics.characteristic: organism
characteristics.text: whole organism
characteristics.characteristic: organism part
characteristics.text: short reads library
characteristics.characteristic: protocol
characteristics.text: Escherichia coli BW25113
characteristics.characteristic: scientific_name
characteristics.text: Sample 2
characteristics.characteristic: title