title: Generic sample from Equus caballus
organism: Equus caballus
description: Equine neurological tissue (cortex, cerebellum, midbrain, hindbrain, thalamus, cervical and lumber spinal cord) and spleen were obtained by necropsy from three groups of two horses each (total of 6 individuals) and consisted of 1) naive horses infected with 1 X 105 WNV, intrathecally 2) non naive horses vaccinated for protection against WNV (Prevenile, Intervet-Schering-Plough) and infected with 1 X 105 WNV , intrathecally and 3) horses that were not infected or vaccinated  Tissues were snap frozen and stored at -80oC until RNA extraction.  Total RNA was extracted using Trizol (Guanidium thiocyanate- Invitrogen, Carlsbad, CA) and a traditional phase separation technique.  Concentration and purity of each RNA sample was assessed using spectrophotometric technology (ND-1000, Nanodrop Technologies, Wilmington, DE) and degradation assessed by microfluidics (Agilent 2100 Bio-analyzer, Santa Clara, CA).  The RNA was converted to a normalized cDNA library (SMART PCR cDNA synthesis kit and Advantage 2 PCR kit, Clontech, Mountainview, CA; QIAquick PCR Purification Kit, Qiagen, Valencia, California; and cDNA Normalization Trimmer Kit, Evrogen, Moscow, Russia).