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In addition, the MSA should be so large that random elimination of sequences from the alignment will not change the amino acid conservation at each site.
After elimination of sequences with ambiguous nucleotides and non-anchor containing sequences, databases were created and individual sequences were queried against the databases with BLAST and DNA-BAR/DEGENBAR.
We cannot rule out the possibility that the criteria for the elimination of sequences containing CpG islands might not be stringent enough, or that other phenomena, linked to transcription or to translation, could induce the observed variations in GC3 content.
Elimination of sequences disturbing alignments and doubles.
Elimination of sequences with divergent residue composition by using the amino-acid composition test from TREE-PUZZLE software [ 35].
Other parameters that were considered include: 1) Elimination of sequences that were identical or almost identical to bacterial sequences.
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Elimination of sequencing reads containing sequencing errors ('technical variants') is usually achieved by matching reads to a finished genome or, since this is not available for barley, through culling reads based on quality information from the sequencer.
This strategy allows for easier elimination of sequence deviations caused by sequencing errors in the sequenced or the reference genomes.
There is a clear danger, therefore, that technical variants of an abundant miRNA in a deep sequencing project can be misidentified as hitherto undiscovered family members and so identification and elimination of sequencing errors is vital to the analysis.
The molecular drive arising from biased DNA uptake would reinforce selection for uptake sequences linked to beneficial alleles and oppose the elimination of uptake sequences associated with deleterious mutations.
MiSeq SAM files of alignment, generated with the GMAP aligner software [ 29] using the human reference genome (hg18/NCBI_36), were further subjected to the elimination of duplicated sequences, known to be prevalently produced during the pre-amplification sequencing step of the Illumina workflow [ 30].
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