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Mutation profiles of 32 mutant samples with 41 mutations were repeated with the ColoCarta.
Selected mutations were repeated by Sanger sequencing and gave identical results to the PGM sequence.
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The initial screening for DNA pools containing mutations was repeated at least three times.
The process of crossover and mutation was repeated until 50 offspring were created.
Mutagenesis was carried out using the BuildModel FoldX command, and each mutation was repeated five times for each structure.
Although no mutation was repeated in all populations, amino acid substitution R55W in non structural protein 2C was present in populations V3B, V19 4B and V15 9B (Table 1).
However, to completely account for the possibility of unreliable SNPs called in these regions, the mutation rate analyses were repeated with these regions removed (Additional file 1).
As before, these conditions were repeated with mutations disallowed, and those results are presented in figure 6.
As more than two-thirds of the mutations are nonsynonymous, these analyses were repeated at the protein level to check the existence of ambiguities.
Mutation profiling was repeated on 32 mutant samples using ColoCarta and the results were identical to results with OncoCarta, demonstrating that this methodology was reproducible.
In one patient who was diagnosed with acute lymphocytic leukemia (ALL), mutation analysis was repeated on DNA isolated from gastric antrum tissue, to confirm the germline status of the mutation.
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