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These three antibodies do not bind to wild-type human and porcine PrP molecules, most likely due to sequence inconsistencies in residues necessary for forming the antigen-antibody bond.
Apparent misassemblies and regions with sequence inconsistencies were corrected in Artemis after PCR and sequencing confirmation of the regions.
Manual inspection of selected SIM4 alignments showed apparent sequence inconsistencies, when compared to the secondary alignments [see Additional File 1].
Sometimes, there are DNA sequence inconsistencies between existing ad hoc genomic reference sequences (where no RefSeqGene record has yet been created) and their NCBI RefSeq mRNA sequence counterparts.
A final assembly assessment was performed by mapping each set of reads to their respective plastome and locating any sequence inconsistencies as described in Wysocki et al. [ 49].
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This finding is unlikely to be due to sequencing inconsistencies, as the methods employed to determine BRAF and KRAS mutation status had been intensively validated [ 28, 38, 39].
In order to monitor sequencing errors that could arise from sequencing, phylogenetic inconsistencies were monitored using phylogenetic tools [ 39- 44].
With data from mixed sources, the sequences for each target node can be aligned in a ranked order; on the other hand, the graph representation of reconstructed sequences tolerates inconsistencies between subsequent alignments and the algorithm can resolve conflicting gap patterns among a set of diverged sequences.
Even if a guide-tree reflects correct phylogenetic relationships, these alignments may be inconsistent with remaining sequences and the inconsistencies are propagated to further steps.
Based on the 2,112 deletions for strain CB4856 retrieved from WormBase WS210 AceDB server, we filtered those with (i) duplicated coordinates, (ii) inconsistency between the reported length of the deletion and the actual sequence, and (iii) inconsistency between the reported deleted sequence and that found in WS210 for the same coordinate.
Comparisons of Illumina and Sanger sequencing revealed two inconsistencies: site 23 in PmaNZ058, and site 12,982 in PmaNZ005 (no longer inconsistent when duplicate Illumina reads were removed), giving a total Illumina-Sanger discrepancy rate of two inconsistencies in 57,240 bp compared or 0.0034%.
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