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Fingerprints of 34,580 BAC clones (5.6× genome coverage) were generated for the FPC assembly of the BAC contigs.
The highest N50 (computed by sorting the contigs from largest to smallest and then determining the minimum set whose sizes total 50% of the assembly) generated for the transcripts is 1,755 generated by a k-mer length of 59. Predicted open reading frames range from 1 to 108,112 bp with the highest number of ORFs being generated with a k-mer of 41.
Importantly, we found that the sequence assemblies generated for the same orthologous regions from various vertebrates show substantial variation with respect to misassemblies and, in particular, the frequency and characteristics of sequence gaps.
Unigenes generated from the assembly were further filtered for a minimum size of 200 nt.
Using this approach, a single contig assembly has been generated for the genome of the wAu strain, which does not induce CI.
An overall summary score was generated for each assembly by combining the number of error-free bases with the scaffold N50 length, calculated before and after breaking the assembly at errors, as follows: Number of error free bases * broken N 50 2 / original N 50 Normalization is also applied within each species (see Methods).
Then, based on the CEP genes identified, general feature format (GFF) files were generated for each assembly which specified CEP mRNA regions as beginning 100 nucleotides upstream of the start codon and 300 nucleotides downstream of the stop codon.
The O. fasciatus and S. vulgaris sequence reads were generated for de novo assembly of the entire transcriptome of the organism while the I. tridecemlineatus sequences were generated for differential expression analysis [ 34– 34].
The 100 bp paired-end reads were generated for transcriptome assembly and 100 bp single-end reads were generated for DGE analysis.
The KEGG maps for more than 130 metabolic pathways were generated for both assemblies and the results were exported.
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