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All the genes not producing a significant alignment with the genome or aligned to unassembled contigs, where the annotation was not available, were classified respectively as "not aligned" and "contig".
Amino acid sequencing of other peptides derived from limited trypsin cleavages and purified by HPLC were included and showed a significant alignment with the recently deduced gastrokine-1 sequence ("Figure 2").
Approximately 17% of the SNPs had flanking sequence data which gave a significant alignment with an annotated gene in the stickleback genome.
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The LG and base pair (bp) location of each gene model producing a statistically significant alignment with the query sequence was compared to the QTL intervals previously projected on the genome.
All 80,357 transcripts were compared with the NCBI non-redundant (nr) protein database using Blastx with an E-value of 1e-10, which resulted in 53,670 (66.8 %) sequences with a significant alignment (Additional file 4).
All sequences that exhibit a significant alignment (E-value of ≤ 7 × 10-13) withethe consensus were retrieved from Unigene http://www.ncbi.nlm.nih.gov/UniGene/UGOrg.cgi TAXID=3635 in the Cotton Genome Database http://cottondb.org/cdbhome.html.html
Thus, if the first significant alignment with a library sequence aligns residues 25 125 at iteration i, later alignment boundaries at iteration i +1 and beyond are ignored; only the initially aligned region (25 125) is used to form the PSSM.
A total of 63,202 contigs (51.3%) with an average length of 1,495 nucleotides had at least one significant alignment with a protein in the non-redundant database of GenBank.
BLAST search of the putative "anticodon arm" returned one significant alignment with a region located between the genes ND3 and ND5 and designated as "Non-translated intergenic spacer" of Steganacarus magnus.
Two genes (TICAM1 and TOLLIP) could not be assigned to a position in the porcine FPC map because a 3 Mb human sequence fragment surrounding the localisation of the genes produced no significant alignment with any porcine clone mapped on the FPC map.
When submitted to protein sequence alignment, this peptide showed only one significant alignment with gastrokine-1 (GKN1).
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