, 2005). Sequences were then assembled into contigs using the OASES sequence assembly software (Schultz et al., 2012). OASES Kmer lengths of between 49 and 59 were evaluated to determine the optimal contig size. Contig ID Protein Tyrosine Kinase inhibitor was determined using a stand-alone BLASTx search against

the Ensembl zebrafish protein database (version Zv8.59, E-value < 1e-10) and contigs that could not be assigned to zebrafish transcripts, splice variants or non-conserved regions of known proteins were eliminated from further analysis. The zebra fish proteome was chosen for identification of contigs despite the fact that databases for species more closely related to barramundi are available (i.e. Takifugu rubripes, Tetraodon nigroviridis), they are not as thoroughly annotated and did not return as high a number of BLASTx matches to known proteins. Sequence reads were then mapped to annotated contigs using Novacraft software ( Li et al., 2009) with count data recorded for each annotated gene

within each sample pool of interest. Weight differences between northern and southern barramundi reared at 36 °C, 28 °C and 22 °C Z-VAD-FMK price were statistically compared by means of ANOVA. Homogeneity of variance was confirmed using a Levene’s test and differences of p < 0.05 between time points were considered significant. All ANOVA testing was performed using SPSS v 16.0 (SPSS, 2006). To detect differentially expressed genes between all four experimental comparisons (N22 vs. N36, N22 vs. S22, N36 vs. S36 and S22 vs. S36) the edgeR package (Robinson et al., 2010) was used in conjunction with R software and customized script commands. Program estimated method of dispersion was generated and applied to the data with a false discovery rate (FDR) cutoff of ≤ 0.05. Gene ontology analysis was then performed upon contigs identified as differentially expressed using the goseq R Bioconductor package (Young et al., 2010) to retrieve

information relating to cellular components, biological processes and molecular functions. Weight data was recorded for both southern and northern barramundi populations reared for ~ 3.5 months (106 days) at 22 °C, 28 °C and 36 °C as a measure of growth and to compare the performance of each population at different temperatures. At a rearing temperature Adenosine of 22 °C southern barramundi showed significantly higher growth after 106 days than northern barramundi (p < 0.001) (Table 1). As expected, at the control rearing temperature of 28 °C there was no significant growth differences between southern and northern barramundi and there were also no recorded growth differences in the final weights of both southern and northern barramundi grown at 36 °C (Table 1.). Within populations, southern barramundi showed significantly higher end weights at 28 °C than at either 22 °C or 36 °C (p < 0.