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The consistent sequence of events leading to fluorescence emission endows fluorophores with several general properties.
For each larger PCR product, at least two independent clones were sequenced to ensure that we obtained the consistent sequence.
This allowed us to compare the effects in response to a violation element in the violation sequences to an acoustically identical element in the consistent sequence (Fig. 1C).
We observed that for a violation element, certain ERP components were modulated to a greater extent than for their acoustically matched elements in the consistent sequence.
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The violation sequences also elicited a significantly greater number of breaches above the confidence intervals than the consistent sequences (Wilcoxon signed-rank test, p < 0.001; mean number of breaches, for violation: 2.85 (±0.18); consistent: 2.07 (±0.16)).
We observed that the distribution of average area breaches across the CI was shifted towards significantly higher areas for the violation, relative to the consistent sequences (Fig. 3C; Wilcoxon signed-rank test, p = 0.013; mean CI breach area (standard error of the mean, SEM), for violation: 12.81, (±1.96); for consistent: 8.73 (±1.17)).
Next each of the EEG waveforms in response to the violation sequences were analysed relative to the EEG responses to the matching consistent sequence pairs.
Critically, the first illegal sound element in a violation sequence was identical to one in the matching consistent sequence, allowing us to analyse acoustically identical parts of the consistent and violation sequences (Suppl. Fig. S3).
The most consistent sequence amplified was 191 bp between mitochondrial nucleotide position (np) 16210 and np16401 (Table 2).
The lack of consistent sequence features precludes a sequence-based recognition.
The most consistent sequences obtained came from the largest fragment investigated (see Additional File 2).
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