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We complemented these tools with sequence alignment, including the DIALIGN and ALIGN-M algorithms designed for highly divergent sequences with only localized similarities, as seen in 5'-NCS.

To investigate in more detail the factors affecting the performance of each program, we characterized each alignment using a number of 'global' attributes describing the overall full-length alignment, including the number of sequences to be aligned, their length, an MSA objective function (norMD) [40] and the percentage of the alignment covered by the blocks.

After the HMM is trained, it is run for every read-half alignment, yielding the coordinates of the first piece of the read alignment, including the first exon-intron boundary of the splice junction.

The final alignment, including the original Pfam set plus our own, additional proteins, will be referred to as the Pfam MSA, even though it is enlarged by a few additional proteins.

A concatenated alignment including the species for which all of the selected ribosomal genes were present was generated.

For phylogenetic analysis, we generated an alignment including the 40 newly generated and 20 additional mtDNA genomes downloaded from Genbank.

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The corresponding forward alignment included the trio.

The alignment included the same amino acid positions that were used for the phylogeny.

Alignment includes the Hox homeodomain, twelve residues 3′ of the homeodomain, and eight residues 5′ of the homeodomain for representative species.

Each alignment includes the genomic sequences of S. cerevisiae as a reference, which is used for annotation of the alignments via known genetic elements from the genome of S. cerevisiae.

This alignment includes the genomes of human (Homo sapiens, version NCBI 36.1, March 2006), chimpanzee (Pan troglodytes, version PanTro 2, March 2006), and rhesus monkey (Macaca mulatta, Baylor College of Medicine Human Genome Sequencing Center, version v.1.0 Mmul_051212, January 2006).

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