CoclBAC 1-5
BAC library construction, screening and clone sequencing of lake whitefish (Coregonus clupeaformis, Salmonidae) towards the elucidation of adaptive species divergence
Using a PCR screening approach, we isolated 10 clones from the lake whitefish BAC library. More specifically, we identified two clones for each of five target genes. Per-target shotgun library preparation, tagging (454 Multiplex Identifiers, MIDs) and sequencing were done at the IBIS biomolecular analysis platform (Université Laval, Québec, Canada) on a 454 Genome Sequencer FLX System, with long-read GS FLX Titanium chemistry.
Genomic DNA sequences and other genomic resources are essential towards the elucidation of the genomic bases of adaptive divergence and reproductive isolation. Here we describe the construction, characterization and screening of a BAC library for lake whitefish (Coregonus clupeaformis). We then show how the combined use of BAC library screening and next-generation sequencing can lead to efficient full-length assembly of candidate genes and their genomic background. The whitefish BAC library represents 5.2 haploid genome equivalents.  Ten BAC clones were isolated following a quantitative real-time PCR screening approach that targeted previously identified candidate genes. Sequencing of these clones on a 454 GS FLX system yielded 178 000 reads with a mean length of 358 bp, for a total of 63.8 Mb. De novo assembly and annotation then allowed retrieval of contigs corresponding to each candidate gene, which also contained up- and/or downstream sequences. These results suggest that the lake whitefish BAC library combined with next-generation sequencing technologies will be key resources to achieve a better understanding of both adaptive divergence and reproductive isolation in the lake whitefish species complex as well as salmonid evolution in general.