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We carefully inspected the alignment to verify its consistency with the literature.
We also manually inspected the alignment (Additional file 1: Figure S1) using IGV [ 19, 20].
We visually inspected the alignment to verify that key sequence motifs (e.g., HRD and DGF motifs) and core hydrophobic residues are aligned correctly.
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We visually inspected the alignments and performed manual refinement.
For some genes of high interest, we manually inspected the alignments of the protein sequences within the orthogroup, which were done with MAFFT (Katoh and Standley, 2013).
To further improve the reference genomes we mapped the RNA-seq data onto the RMP genomes and visually inspected the alignments using the Artemis Comparison Tool (ACT), a genome viewing tool [ 35].
In addition, we did not validate the variants by Sanger sequence but rather inspected the alignments of short reads using IGV, a method that we have found more sensitive than Sanger sequencing.
To test this possibility, we aligned all the conseqs with this condition (a total of 1,389 clusters representing 329 genes) and visually inspected the alignments to determine why the sequences had not been combined into the same cluster if they were homologous to the same region in D. melanogaster.
During the process we keep track of the inliers rate within the set C ab, as well as the time required by each technique to determine their best correspondence triplet within C ab. Once the alignments are terminated, we visually inspect the alignment outcome and count the number of errors for each technique, reported in Table 1.
In the remaining cases both alleles were observed by inspecting the alignment manually, but Maq failed to call both of the SNP alleles.
The leader sequences, framework regions, complementarity determining regions (CDRs) and J-regions were identified by inspecting the alignment of the F1-40 heand and light chains to other antibody sequences [26] [30].
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