High efficiency ligations capture synthetic miRNAs with high fidelity. (A) Deep sequencing data of an equimolar mixture of 29 synthetic miRNAs from high efficiency ligation reaction (blue bars) and previous method (red bars); the dashed line indicates expected result. For all high efficiency ligations, plotted is the average of at least two independent experiments where error bars represent the standard error of the mean. (B) As in (A) except the mix is composed of an equimolar mixture of 25 synthetic miRNAs (blue bars) and differential mixture of 4 let-7 miRNAs (black bars) where the ratio was let-7i:let-7c:let7f:let-7d = 0.01:0.1:1:10. (C) As in (A) except the synthetic mixture was combined with 0.5 μg C. elegans total RNA prior to ligations.