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hed in homologs of sequences transcribed in mouse, The finding NSC 14613 that, contrary to the situa tion observed with contigs, more singletons had hits to genome Ferrostatin-1 than to EMCT is consistent with the well known fact that the expression level of most noncoding genome transcripts is generally low and tissue or even cell type specific, This may also explain the lack of reports of noncoding transcripts in the previous 454 studies of tran scriptomes in nonmodel organisms. Either coverage was not sufficient in those studies, or the lack of a moderately divergent model organism, enabling meaningful nucle otide nucleotide similarity searches against the genome, precluded the identification of noncoding transcripts.

Certainly, further experimental studies involving RT PCR or microarrays would be necessary to validate further our hypothesis and provide more decisive answers as to whether noncoding RNAs indeed represent SKI II a substantial portion of the bank vole normalized heart cDNA library. SNP differences between selection lines We identified over 1,000 of putative SNPs that showed apparently significant frequency differences between lines. These polymorphisms constitute an abundant source of candidates for genes underlying microevolu tionary response to selection on increased maximum metabolic rate. Overrepresentation of mitochondrial genes among those with SNP frequencies differentiated between selection regimes may be an artifact resulting from generally high coverage of transcripts for mitochon drial proteins in our data.

The candidates will be further validated and investigated using methods allow ing large scale SNP genotyping on an individual basis, The search for Resonance (chemistry) genes underlying the response to selection will be facilitated by construction of a genetic map, which has not yet been developed for the bank AZD3514 vole. Single nucleotide polymorphisms and micro satellite markers identified in this study will be useful for this purpose. Conclusions In the present paper, we report the first comprehensive sequence analysis of the bank vole transcriptome. The heart transcriptome was sequenced in the lines selected for high metabolism and in control lines. Longer reads and higher sequence yield per run provided by the 454 Titanium technology proved beneficial for the assembly quality. We detected transcripts of over 14,000 genes, and, for a substantial fraction of them, the full length of coding regions were obtained.

Almost full representation NSC 14613 of genes known to be expressed in the mouse heart was identified. In addition to genes from the mouse ENSEMBL collection, patterns observed in our data were consistent with widespread transcription from noncod ing genomic regions, a finding not reported in previous studies about transcriptomes in non model organisms. We also detected a number of putative SNPs. a much higher fraction of SNPs than expected by chance exhib ited variant frequency differences between selection regimes. These SNPs are thus promising candidates for causal genetic factors underlying response to selection on AZD3514 metabolic rate.

The transcript sequences generated in the present study constitute a valuable permanent resource forming a foundation for RNAseq experiments aiming in detection adaptive changes both at the level of gene expression and sequence variants, that would facilitate studies of the genetic basis of evolutionary divergence. Methods cDNA preparation NSC 14613 and 454 sequencing Four lines selected AZD3514 for a high metabolic rate and four unselected, control lineages were used in the experiment, The experimental design and measurement protocols followed internation ally recognized guidelines for the research on animals, and were approved by the I Local Ethical Committee for Experiments on Animals in Kraków, according to Polish State Law, M1ACGG was used instead of the M1 primer recom mended by the TRIMMER manufacturer, so that it did not anneal to the 5 end of the first strand cDNA contain ing disrupted polyT sequence. Only polTM1 annealed to this