Mutation spectra of <i>can1</i> mutants generated on canavanine plates exhibit several significant differences with those generated in culture.

<p>(A) The overall distribution of different <i>can1</i> mutation categories is not altered between pre-plating(large) and post-plating (small) <i>can1</i> mutants. The “other” category contains multiple mutations per <i>can1</i> ORF, usually clustered together. (B) Pre-plating (large) but not post-plating (small) <i>can1</i> mutants contained transcription-associated mutation (TAM) hotspots. Lippert et al identified the five main TAM hotspots in <i>CAN1</i>, with ATΔ at CATAT at position 1127 being the most frequent in both their study and ours <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003680#pgen.1003680-Lippert1" target="_blank">[39]</a>. (C) Post-plating mutants were characterized by a significant increase in single nucleotide deletions (predominantly occurring in mononucleotide runs; <a href="http://www.plosgenetics.org/article/info:doi/10.1371/journal.pgen.1003680#pgen.1003680.s005" target="_blank">Table S1</a>). (D) Base pair substitution spectra differed significantly between pre- and post-plating mutants.