title: GSM4658993: Tn-Seq_Caulobacter_crescentus_NA1000(tipFpSA607); Caulobacter vibrioides NA1000; OTHER
library_construction_protocol: For each collection, all resistant clones were pooled and chromosomal DNA was extracted using DNAzol (Lucerna-Chem AG, Switzerland). Purified genomic DNA was used to generate barcoded Tn-Seq libraries and submitted to Illumina NextSeq High sequencing (Fasteris SA). Tn insertion-specific reads (50 bp long) were sequenced using the himar-Tnseq primer (5'- AGACCGGGGACTTATCAGCCAACCTGTTA-3') and then mapped (Map_with_Bowtie_for_Illumina_V1.1.2) to the C. crescentus NA1000 genome (NC_011916.1) using the web-based analysis platform Galaxy (https://usegalaxy.org). Using Samtool_V0.1.18, BED file format encompassing the Tn insertion coordinates were generated and then imported into SeqMonk V1.40.0 (www.bioinformatics.babraham.ac.uk/projects/) to assess the total number of Tn insertion per chromosome position (Tn-insertion per millions of reads count) or per coding sequence (CDS truncated from the first 50 and last 50 bp). SeqMonk datasets were exported into Microsoft Excel files for further analyses.
attributes.tag: GEO Accession
attributes.value: GSM4658993