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Prior to alignment of the concatenated sequence, the sequence of each housekeeping gene was aligned separately and trimmed where necessary.
Clusters were also filtered to identify likely informative families prior to alignment and tree-making.
Reverse sequences were reverse complemented (http://bioinformatics.org/sms/rev_comp.html) prior to alignment using ClustalW.
Prior to alignment, contaminant sequence reads exactly matching the 33bp Illumina GA adaptor sequence were removed using the MAQ program.
As such, a separate synteny mapping algorithm must be applied to map collinear homologous segments among two or more genomes prior to alignment.
This potentially gives an advantage to MLAGAN, TBA, PECAN, and MAVID in the accuracy comparison, as in many cases of biological interest, a reasonable guide tree may not be identifiable prior to alignment.
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Electropherogram from all sequences were checked for quality control prior to alignments using the bioedit software (Hall, Carlsbad, CA, USA).
The promer module was used to generate alignments at the protein level by translating the genome in all six reading frames prior to alignments.
Hence, in Adaptive GDDA-BLAST, we align the query and the target sequence first, and then compute the seed-inserting positions from the alignment result, prior to aligning the chimera sequences.
With the improvement in alignment quality and quantity seen here, we recommend trimming prior to any alignment and downstream analysis, especially for poor quality data.
Although the common interspersed repeats were already removed prior to the alignment, low copy repeats or segmental duplication may still cause mis-alignments.
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