Mf min is the proportion of residues that CHMFL-ABL-053 includes different mutations in the set of sequences (reads), counting only once the mutations repeated at the same genomic position in different clones (Equation I; equations are given in section “Equations and practical examples”). Mf max is the proportion of residues that include mutations in the set of sequences (Equation IIb). Mutations are counted by comparison of each individual sequence with the consensus sequence of the corresponding population. The latter is either determined experimentally by direct sequencing of a PCR product or built with the most frequent nucleotide at each genomic position in the aligned sequences. Sets of compared sequences from different mutant spectra can have different numbers and lengths since the division by the total number of nucleotides sequenced introduces the required normalization (Equations I and II). It is advisable, however, that a similar and sufficient number of sequences be compared to minimize statistical biases.
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