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After screening the reads through alignment with reference cp genomes and extensive contig extension efforts (see Materials and Methods for details), we collected 369,022 cp-genome-related reads (8.8% of total reads and an average of 384 bp in length), reaching 1,081x coverage on average over the cp genome.
The presence or absence of the established NAI resistance-associated mutations, R292K and E119V, was then determined by inspection of the resulting amino acid alignment, with reference to the influenza A/Syd/5/97 (H3NA) NA sequence, which was isolated before the licensing and widespread use of neuraminidase inhibitors that began after 1999/2000.
Alignment with reference sequences and phylogenetic analyses were conducted by using MEGA 5.0 (5.0.
In BAC 604D2, six sequence contigs could be arranged through identification of target site duplications and alignment with reference transposon sequences whereas in BAC 509D2, a CACTA transposon was used to connect two large sequence contigs.
Single-nucleotide polymorphisms (SNPs) were determined by alignment with reference strains (M. tuberculosis H37Rv accession no. NC_000962.3 or M. canettii cluster A Percy3 [STB-D CIPT140060008 accession no. NC_019950.1]) as described (7 ; Technical Appendix 1).
Positions of substitutions are described as amino-acid locations inferred from an alignment with reference sequences of Mus musculus acquired from the NCBI RefSeq protein database [Genbank: YP001686700 for CO1, NP904331 for CO2].
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The details of BLAST outputs including BLAST scores and the sequence alignments with reference are also shown in the report.
For all isolates, the tuf gene was sequenced with species determination based on alignments with reference isolates.
To assess the quality of FlexSnap alignment compared to other structural alignment methods, we evaluated the agreement of the methods' alignments with reference manually-curated alignments.
Our approach was based on the premise that filtering uncharacterized proteins on the basis of their global sequence similarities with the "Reference set" proteins does not reduce their probability of having small/local alignments with "Reference set" proteins.
The web site also allows to perform alignments with reference genomes using on the fly BLAT search against the latest releases of the zebrafish and stickleback genomes in the UCSC Genome Browser.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com