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At least two runs of the M0 model were performed on each alignment to check the consistency of the log-likelihood values between the multiple runs.
Several annotated sequences were then used for an additional alignment to check for conservation of H104.
Two runs of the M0 model were performed on each alignment to check that values for log-likelihood, κ and branch lengths were consistent between the two runs.
Clean-up of sample sets involved preliminary alignment to check for sequence instability or misalignments, as well as confirmation of gene annotation by comparative analyses.
We used multiple alignment to check whether an amino acid that is deleterious in flies can co-occur with D. melanogaster amino acids at this interacting sites.
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The phylogenies of single-gene families were reconstructed based on nucleotide and amino acid sequence alignments (to check for congruence) in a Bayesian Markov chain Monte Carlo (MCMC) statistical framework using MrBayes 3.2 (Ronquist et al. 2012), as well as a maximum likelihood (ML) method implemented in RAxML 7.7 (Stamatakis 2006).
This precision task requires the exact manual positioning of the tape and a careful visual control in order to continuously adjust alignment and to check for any air bubbles fastened under the tape.
We analyzed the synteny of the structured RNA annotation using the pairwise alignments is to check the coverage of a given annotation by transferring the coordinates to the target species using the pairwise alignment (liftOver).
The DNA sequences were aligned using ClustalW v. 1.4 [ 62], and resulting alignment was translated to check for the presence of stop codons.
We only retained the best match for each contig, and the resulting alignment was parsed to check for mismatches, insertions and deletions.
We then generated a reverse complement sequence from the entire set, and by aligning each sequence in a pair-wise alignment to HXB2, checked the similarity so that we kept only the forward direction version of the sequence.
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