organism: Ovis aries
attributes.tag: Age
attributes.value: 190 days
attributes.tag: Alias
attributes.value: E-MTAB-445:Gastric LN_130
attributes.tag: Broker name
attributes.value: ArrayExpress
attributes.tag: Description
attributes.value: Protocols: Uninfected control lambs, were sham dosed with water at the same time as three times a week starting at approximately 12 weeks of age and continuing until sacrifice at approximately 27 weeks. Gastric lymph nodes were collected from all lambs, diced and placed in RNAlater (Ambion) at 20C, incubated at 4C overnight and stored at -70C. Total RNA was isolated from tissues using the Ambion Ribopure (http://www.ambion.com/techlib/prot/fm_1924.pdf). Approximately 100 mg tissue was thawed to 20C and excess RNALater removed by blotting. 2 ml TRI Reagent was added and tissues homogenized using a rotor-stator homogenizer. Insoluble material was removed by centrifugation at 12,000 x g. 1 ml supernatant from each sample was dispensed into 1.5 ml microcentrifuge tubes with 100 ul of bromochloropropane (BCP). Tubes were vortexed and incubated at 20C for 15 min. Tubes were centrifuged at 12,000 x g for 10 min at 4C. 400 ul of the aqueous phase (top layer) was transferred to a new 1.5 ml microcentrifuge tube with 200 ul 100% ethanol, vortexed for 5 s. Each sample was transferred to a Filter Cartridge-Collection Tube assembly, centrifuged at 12,000 x g for 30 s at 20C. 500 ul of Wash Solution was added to the Filter Cartridge-Collection Tube centrifuged for 30 s at 20C and rewashed. RNA was eluted by the addition of 100 ul Elution Buffer incubated at 20C for 2 min and centrifuged for 30 s to elute the RNA from the filter. RNA quality and integrity was assessed using a RNA 6000 Nano LabChip on the Agilent 2100 Bioanalyzer and quantified using a NanoDrop ND-1000 spectrophotometer.
attributes.tag: DevelopmentalStage
attributes.value: lamb
attributes.tag: INSDC center alias
attributes.value: ROSLIN
attributes.tag: INSDC center name
attributes.value: The Roslin Institute and Royal (Dick) School of Veterinary Studies, University of Edinburgh
attributes.tag: INSDC first public
attributes.value: 2010-12-07T15:35:34Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T15:25:17Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: OrganismPart
attributes.value: gastric lymph node
attributes.tag: SRA accession
attributes.value: ERS017440
attributes.tag: Sample Name
attributes.value: ERS017440
attributes.tag: Sex
attributes.value: female
attributes.tag: Title
attributes.value: Gastric LN_130