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For O. rufipogon, all reads aligning to mPing, Ping, and Pong were pooled to analyze the base composition at the + 16 G/A SNP because mPing, Ping, and Pong insertions could not be efficiently sorted.
We did not use occurrence predictions that assume a different base composition at the 1st-, 2nd-, and 3rd-codon position, predictions that albeit attractive in terms of fitting potential are biologically absurd since they require a mechanism that lets DNA sequences mutate differently at the three codon positions.
In addition, the base composition at the third codon position for some genes is quite different from the other positions.
Other websites such as the OGRe database of mitochondrial genomes [ 31] also allow analysis and graphing of base composition at the codon positions as well.
13– 15 The characteristics of codon usage are strongly correlated with the genomic GC content, especially the base composition at the third codon positions.
Following DNA phosphatase treatment the base composition at the genomic position preceding the sequencing start was found to be enriched in pyrimidines (position −1; Figure 4, middle).
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Base composition, AT-skew and codon usage were calculated for the eight newly sequenced genomes and the 12 published Bombycidae mitochondrial genomes (Table 1).
This dependence of codon usage on genome base composition (AT- or GC-richness) has been previously reported in several unicellular genomes.
As a consequence, these unfavorable features result in difficulties in under-represented genome regions, particularly when the sequences are from genomes with base compositions at the extremes of high or low G+C content.
The base composition at each of the three codon positions varies, with the first position having the lowest proportion of A+T and the third position the highest (Table 1).
The analyses of the base composition at each codon position of the concatenated 13 PCGs of P. citri show that the third codon positions (92.9%) have an A + T content higher than that of the first (82.8%) and second (79.4%) codon positions, as has also been found in other sequenced Acari (Additional file 4).
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