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Sequencing Salmon for Evolution's Secret

GENERAL - With a whole genome duplication event and wealth of biological data, salmonids are excellent model organisms for studying evolutionary processes, fates of duplicated genes and genetic and physiological processes associated with complex behavioral phenotypes.

According to BMC Genomics 20, it is surprising that no salmonid genome has been sequenced.

Atlantic salmon (Salmo salar) is a good representative salmonid for sequencing given its importance in aquaculture and the genomic resources available. However, the size and complexity of the genome combined with the lack of a sequenced reference genome from a closely related fish makes assembly challenging.

Given the cost and time limitations of Sanger sequencing as well as recent improvements to next generation sequencing technologies, Sanger examined the feasibility of using the Genome Sequencer (GS) FLX pyrosequencing system to obtain the sequence of a salmonid genome. Eight pooled BACs belonging to a minimum tiling path covering ~1 Mb of the Atlantic salmon genome were sequenced by GS FLX shotgun and Long Paired End sequencing and compared with a ninth BAC sequenced by Sanger sequencing of a shotgun library.

The results of this study show that an initial assembly using only GS FLX shotgun sequences (average read length 248.5 bp) with ~30x coverage allowed gene identification, but was incomplete even when 126 Sanger-generated BAC-end sequences (~0.09x coverage) were incorporated. The addition of paired end sequencing reads (additional ~26x coverage) produced a final assembly comprising 175 contigs assembled into four scaffolds with 171 gaps.

Sanger sequencing of the ninth BAC (~10.5x coverage) produced nine contigs and two scaffolds. The number of scaffolds produced by the GS FLX assembly was comparable to Sanger-generated sequencing; however, the number of gaps was much higher in the GS FLX assembly.

The team concluded that these results represent the first use of GS FLX paired end reads for de novo sequence assembly. Our data demonstrated that this improved the GS FLX assemblies; however, with respect to de novo sequencing of complex genomes, the GS FLX technology is limited to gene mining and establishing a set of ordered sequence contigs.

Ellen Hardy

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