Figure 2

Sequence coverage from direct MiSeq PE250 sequencing. A: Direct sequencing of target amplicons without shearing produces homogeneous sequence coverage. Representative average read coverage of three amplicons under a single barcode with high (blue), medium (red) and low (green) coverage. Even the poorest sequence coverage exceeds the minimum coverage needed to accurately identify mutations in our 288-fish pools (red line). B: Amplicon coverage across all library pools. Different colors represent the same amplicon from each of the 32 pools in the library. C: Variable coverage of different fragments. Each bar represents the average coverage across the entire library of a single randomly selected nucleotide in each of 109 fragments screened. 107 of 109 fragments exceeded the minimum coverage needed to accurately identify mutations in our 288-fish pools (red line).