Project description:Sheep (N=6) infested with Psoroptes ovis mites were bled weekly and circulating leukocytes isolated from their whole blood samples. RNA was extracted and processed onto Agilent Bovine Microarrays and differentially expressed genes identified between the following time points (0 (Baseline (pre-infestation)), 1 week, 3 weeks and 6 weeks post-infestation).
Project description:Transcriptional analysis of ovine skin response to infection with the parasitic mite Psoroptes ovis using the Agilent ovine transcriptome microarray platform. The results of statistical analysis (differential gene expression across the time course of infection was determined using a one way-analysis of variance (ANOVA) with a Student-Newman-Keuls (SNK) post-hoc test in Genespring GX 11.0 (Agilent Technologies, UK) comparing each of the 5 time points, non-infected and 1, 3, 6 and 24 hours post infection. Multiple test correction was performed using the Benjamini & Hochberg False Discovery Rate (FDR) procedure with an FDR corrected p-value cut-off set at 0.05) and fold change analysis (Fold change analysis was performed on the one-way anova dataset, probes with a fold change greater than 2 between any of the conditions were carried forward) are in archive E-TABM-1012.additional.zip. Quality control: Six biological replicates (sheep, n=6), multiple biopsies pooled from each animal at each time point of infection. Pooled samples from each sheep at each time point hybridised in single-dye (Cy3) format to Agilent ovine arrays. Positive and negative control probes utilised to assess array QC and real time qRT-PCR validation of selected array probes