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Similar to previous results in human cell lines (Komor et al., 2016), besides C→T conversion, we observed rare C→G substitution (sample #21) and C→A conversion (sample #22).
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The objective of this study was to investigate substitution sampling as an alternative recruitment strategy.
Apart from this single substitution, samples from all animals of herd #17 were identical, including the three sequences obtained from samples prior to disease outbreak in herd #17.
Moreover, the average loss factor and the loss factor under 1 T of the sample (flower-like CIPs weight content 8 wt%) are 0.36 and 0.07, which are approximately 1.71 and 2.71 times than that in the non-substitution sample.
We further assessed the contributions of substitutions sampling error and showed that the observed variance greatly exceeds the variance that can be attributed to sampling.
The G12A substitution in sample 11 was present in 21% of 1358 reads and the T790M resistance mutation in sample 10 occurred at a frequency of 20% out of 909 reads (Table 1).
One sequence from a tick fed on a raccoon had 1 additional substitution; this sample had a melting point maximum of 82.5°C.
(B ) Average number of somatic substitutions per sample across 31 tumor types.
After water in form of ice within the cells was replaced by substitution media, samples were embedded in epoxy resin.
For freeze substitution, tissue samples were immersed in a solution of 0.5% uranyl acetate in anhydrous methanol and kept overnight at −90 °C.
Divergence times were estimated using a relaxed-clock method with substitution rates sampled from an uncorrelated lognormal distribution implemented in BEAST v. 2.1.3 [ 37].
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