organism: Ovis aries
description: Protocols: New Zealand Cheviot sheep of PRNP genotypes VRQ/VRQ, from the DEFRA breeding flock were inoculated subcutaneously with 2 ml of 10% w/v brain homogenate into sites on either side of the neck roughly 10cm anterior to the prescapular lymph node (PSLN). In all experiments frozen lymph node was converted to a frozen tissue powder. Briefly a 100 mg of tissue per sample was placed into a Teflon flask along with steel ball and placed in liquid Nitrogen for 1 min. Homogenised for 60 seconds at 2000 rpm in a Mikro-Dismembrator U (B.Braun Biotech International GmbH). Total RNA was extracted from the frozen tissue powder using the RiboPure Kit (Ambion) including a DNase I digestion. To the frozen tissue powder 1 ml of Tri Reagent was added and then homogenise for a further 15 seconds at 1500rpm. A 200 ul aliquot of chloroform was added to lysate and mixed vigorously by vortexing. The protocol was then followed as per the manufactures instructions. The RNA was eluted in a final volume of 50 ul. Each RNA sample was DNase I (Ambion) treated as per the manufacturers instructions. The quality and quantity of the isolated total RNA was assessed using a RNA LabChip on the Agilent 2100 bioanalyser and a NanoDrop ND-1000 Spectrophotometer (Nanodrop Technologies Inc.). (
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2014-06-01T17:02:25Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-03-08T17:15:54Z
attributes.tag: External Id
attributes.value: SAMEA2397564
attributes.tag: INSDC center name
attributes.value: University of Edinburgh
attributes.tag: INSDC first public
attributes.value: 2014-06-01T17:02:25Z
attributes.tag: INSDC last update
attributes.value: 2018-03-08T17:15:54Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-2385:OB_K331
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: common name
attributes.value: sheep
attributes.tag: developmental stage
attributes.value: lamb
attributes.tag: disease
attributes.value: standard scrapie sheep prion (SSBP/1) infected
attributes.tag: organism part
attributes.value: Obex
attributes.tag: sample name
attributes.value: E-MTAB-2385:OB_K331
attributes.tag: scientific_name
attributes.value: Ovis aries
attributes.tag: sex
attributes.value: male
attributes.tag: specimen with known storage state
attributes.value: frozen specimen