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In fact, the number of orthologs did not increase when decreasing the minimum required sequence identity levels to as low as 50%% in any gene family (data not shown).
Additional file 1, Table S4 gives the numbers of singletons obtained when varying the minimum required sequence identity and coverage of the blastp hits in the range from 70-10070-100%
Exonerate (Slater and Birney 2005) was used with the parameters "exhaustive" (which turns all heuristics off and allows for the most sensitive search), "percent10" (which sets the required sequence identity threshold for putative hits to 10%), and "est2genome" (which models cDNA to genome alignments and allows for alignments to be interrupted by nonconserved regions such as introns).
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Hits were filtered to require sequence identity > = 90%, and an alignment length > = 64.
Using a stringent cutoff requiring sequence identity of at least 90% and a minimum alignment length of 30 nucleotides, 509 (38.6%) sequences in our library have a hit in the previous library, and 711 (54%) of the sequences in the earlier library have a hit in ours.
Given the limited number of known RNA structures, structural motif templates with the required high sequence identity are difficult to attain.
When aligning to the transcriptome, we required 98% sequence identity over more than 30 bases, with no intronic gaps, '-minIdentity = 98 -minScore = 30 -maxIntron = 0'.
For example, Dynalign (Mathews and Turner 2002) combines free energy minimization and comparative sequence analysis to find a low free energy structure common to two sequences without requiring any sequence identity.
We maintain that evolutionarily related targets logically provide molecular avatars, where this Sanskrit term for descent includes ideas of functional relationships and of being a physical embodiment of the target's essential features without requiring high sequence identity.
Our analysis requires ≥92% sequence identity during contig assembly (Huang et al. 2003).
For commercial applications that require the sequence identity to be confidential, we have an encryption schema for nuID.
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