Fraction of probes above an arbitrarily defined threshold of 1750 fluorescence units.

<p>(<b>A</b>) Probes designed to have a melting point of 18–22% formamide and hybridized at 0% (red), 15% (green), and 20% (blue) formamide. Left panel, <i>E. coli</i> probes hybridized with 5 ng <i>E. coli</i> target (PM data) and 50 ng <i>R. sphaeroides</i> non-target (MM data); right panel, <i>R. sphaeroides</i> probes hybridized with 50 ng <i>R. sphaeroides</i> target (PM) and 5 ng <i>E. coli</i> non-target (MM). (<b>B</b>) The predictive power of hybridization efficiency for <i>E. coli</i> probes hybridized with 50 ng <i>R. sphaeroides</i> (dashed lines) and <i>R. sphareoides</i> probes hybridized with 5 ng <i>E.coli</i> (solid lines) for all mismatches (red), 1–2 mismatches (green), and 3–5 mismatches (blue). Data from formamide concentrations 10, 15, 20, and 25% were combined to maximize the sample space for each data point. x-axis shows midpoints of bins with a hybridization efficiency window of 0.1, except for end bins (window of 0.05).