characteristics.text: ERC000011
characteristics.characteristic: ENA-CHECKLIST
characteristics.text: 2013-10-15T17:00:43Z
characteristics.characteristic: ENA-FIRST-PUBLIC
characteristics.text: 2018-03-08T16:43:28Z
characteristics.characteristic: ENA-LAST-UPDATE
characteristics.text: SAMEA2178380
characteristics.characteristic: External Id
characteristics.text: Institut Pasteur
characteristics.characteristic: INSDC center name
characteristics.text: 2013-10-15T17:00:43Z
characteristics.characteristic: INSDC first public
characteristics.text: 2018-03-08T16:43:28Z
characteristics.characteristic: INSDC last update
characteristics.text: public
characteristics.characteristic: INSDC status
characteristics.text: ERS346489
characteristics.characteristic: SRA accession
characteristics.text: E-MTAB-1877:cell_culture_1
characteristics.characteristic: Submitter Id
characteristics.text: ArrayExpress
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0009737
characteristics.characteristic: broker name
characteristics.text: house mouse
characteristics.characteristic: common name
characteristics.text: Protocols: All experiments involving mice were handled in accordance with the Pasteur Institute guidelines for animal welfare. Only 9- to 12-week-old female C57BL6/J conventional (Charles River) and germfree wild type mice mice (CDTA) were used for experiments. Germ-free mice were housed in plastic gnotobiotic isolators. L. monocytogenes overnight cultures were diluted in BHI and bacteria were grown until OD600nm=1. Bacterial cultures were centrifuged at 3,500 x g for 15 min. After three washes in PBS, L. monocytogenes pellet was resuspended in PBS at a final concentration of 2.5 x 1010 bacteria/mL. Mice were infected orally with 5 x 109 bacteria diluted in 200 uL of PBS supplemented with 300 uL of CaCO3 (50 mg/mL) for 24 h and 72h. Serial dilutions of the inoculum were plated to control the number of L. monocytogenes inoculated in mice. RNAs from the ileal tissue were extracted and purified using classical TRIzol/chloroform protocol. All samples were treated with Turbo DNase (Ambion). RNA quality was determined using Experion Automated Electrophoresis Station (Bio-Rad). The purified small RNA was ligated with 3' RNA adaptor (5'-/5rApp/ATCTCGTATGCCGTCTTCTGCTTG/3ddC/), which is specifically modified to target miRNAs and other small RNAs that have a 3'hydroxyl group. The 5' RNA adaptor (5'-GUUCAGAGUUCUACAGUCCGACGAUC) was then ligated to the 5' phosphate end of small RNA. Reverse transcription was done to convert the RNA to cDNA, which was then selectively enriched by 12 cycles of PCR. The PCR products were purified on 5% PAGE (BioRad) in a size range between 90-105 base pairs and checked on a Bioanalyser DNA1000 chip (Agilent).
characteristics.characteristic: description
characteristics.text: conventional
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/CL_0000624
characteristics.characteristic: growth condition
characteristics.text: uninfected
characteristics.ontologyTerms: http://www.ebi.ac.uk/efo/EFO_0001460
characteristics.characteristic: infect
characteristics.text: Mus musculus
characteristics.characteristic: organism
characteristics.text: Mus musculus
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/NCBITaxon_10090
characteristics.characteristic: organism
characteristics.text: ileum
characteristics.characteristic: organism part
characteristics.text: Mus musculus
characteristics.characteristic: scientific_name
characteristics.text: fresh specimen
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/OBI_0000971
characteristics.characteristic: specimen with known storage state
characteristics.text: 0 hour
characteristics.ontologyTerms: http://purl.obolibrary.org/obo/UO_0000032
characteristics.characteristic: time
characteristics.text: cell_culture_1
characteristics.characteristic: title