characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2022-03-09T00:13:56Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2022-03-09T00:13:56Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA10113883
characteristics.characteristic: External Id
characteristics.text: Institute for Integrative Biology of the Cell (I2BC), Universite Paris-Saclay, CEA, CNRS, Universite Paris Sud
characteristics.characteristic: INSDC center name
characteristics.text: 2022-03-09T00:13:56Z
characteristics.characteristic: INSDC first public
characteristics.text: 2022-03-09T00:13:56Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS7770437
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-10951:FX482
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: Protocols: Integration assays: V. cholerae recipients (CMV30 and EPV369) were grown to an OD600 of 0.3 in LB with 0.2% of L-arabinose (L-ara), to induce the Xer machinery. E. coli β2163 donors were grown to an OD600 of 0.6 in LB supplemented with 0.3 mM of Dap, and mixed with recipients at a 1:10 ratio. Donor and recipient cells were incubated for 3h on LB agar plates supplemented with Dap and L-ara, resuspended in LB supplemented with 0.2% of L-ara and incubated for an additional 1h. Conjugants were selected for the plasmid antibiotic resistance (chloramphenicol)and Dap autotrophy. In the case of the degenerate plasmid libraries, several conjugations were performed for each integration assay to ensure the recovery of ~10+6 clones. Stability assays: Integration libraries were incubated without any selection pressure for the maintenance of the integrated elements. Fresh LB was inoculated with ~1x10+9 cells from the integration libraries, with the addition of arabinose 0.2%. genomic DNA extraction kit Plasmid attP sites were amplified by performing 17 PCR cycles on 50-100 ng of plasmid library gDNA with an equimolar mix of 4103-4105 P5 and 4178-4180/4246-4248 P7 primers. The products were purified from the P5 and P7 primers the double selection with AMPure.
characteristics.characteristic: description
characteristics.text: XafT+ ; n8gtg
characteristics.characteristic: mini-TLC genotype
characteristics.text: Vibrio cholerae
characteristics.characteristic: organism
characteristics.text: Vibrio cholerae
characteristics.characteristic: organism
characteristics.text: Vibrio cholerae
characteristics.characteristic: scientific_name
characteristics.text: dif1 at lacZ locus
characteristics.characteristic: strain genotype
characteristics.text: FX482
characteristics.characteristic: title