Figure 3

The quality of RNA extracted from fathead minnow tissues using five commercially available RNA isolation kits. The panels on the left (A, C, E, G and I) show the 260/280 ratios of RNA extracted using each kit; the dashed lines on each panel represents a 260/280 ratio of 1.8 (the value above which RNA is considered to be "pure"). The panels on the right (B, D, F, H and J) show the RNA integrity number (RIN); the dashed line represents a RIN of 5 (the minimum acceptable RIN for qPCR analysis), while the dotted line shows a RIN of 7 (the minimum recommended RIN for microarray and next generation sequencing analyses). Reported p-values were obtained by one-way ANOVA, except for those denoted by a "W". The "W" indicates that significant differences in variance were detected and that analysis was conducted via a Welch ANOVA. Error bars indicate standard deviation. Different letters above bars indicate significant differences in the measured RNA quantity parameter between samples processed by each of the kits. Percentages reported in each bar are indicative of the coefficient of variance. RIN values were not measurable (indicated by "nm") in RNA extracted from embryos using the SimplyRNA HT kit due to low RNA concentrations.