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Direct sequencing yielded an unambiguous sequence without evidence of mixed sequences in 7 specimens.
Short clean reads were loaded into computer and the unambiguous sequence fragments as contigs were output.
Population genetic analysis was conducted for a partial fragment of the gene encompassing nucleotide coordinates 87-2862 (unambiguousequencece available for 96 isolates).
In contrast, pyrosequencing provides unambiguous sequence from the first nucleotide 3' to the primer, increasing the sequence information yield per assay.
Of the 18 positive PCRs, 14 produced unambiguous sequence data, and were identical to each other across the homologous 943 nucleotide (nt) region sequenced, with the remaining four being bacterial mixtures of indiscernible species composition.
Short clean reads were loaded into computer and the unambiguous sequence fragments were obtained as contigs.
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Although the predicted S. gregaria precursor displays putative hallmarks of an NPLP precursor, comparative sequence analyses of NPLP precursors have revealed substantial heterogeneity among different species, impeding unambiguous sequence-based identification of NPLP orthologs.
The list of specimens, for which we succeeded in receiving amplifyable DNA and unambiguous sequences is given in Table S1.
Following enzymatic purification with ExoSAP-IT™ (Usb), sequence reactions were performed using primer Phe1R, which allowed to obtain unambiguous sequences of the entire control region.
However, Consortium for the Barcoding of Life (CBOL) disregarded trnH-psbA as it does not consistently provide bidirectional unambiguous sequencing reads [17].
Among almost 20 proteins identified by unambiguous sequencing of at least two peptides (Figure 1C; Table 1) we found histones of all the subfamilies, namely H1, H2A, H2B, H3 and H4.
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