Simulated cumulative frequencies of all detectable single base sequence variation events in the 362 amplicons of the E. coli-K-12 reference set. 97%–100% of all possible deletions are detectable in a total of 319 amplicons, 97%–100% of all possible insertions are detectable in 314 amplicons, and 97%–100% of all single nucleotide substitutions can be detected in 355 amplicons. (Black) Total single base variation events; (gray) SNPs, insertions, and deletions.