Fig. 5

Post-UNG bead-based purifications: library yield data. a Effect of bead to DNA ratio on library sizes. Various combinations of 1:1 and 2:1 bead to DNA ratio were applied for post-ligation and post-UNG purifications. The final purified PCR product was run on Agilent DNA 1000 for size profiling. b Size profiles of libraries made using variations of the UNG step. The first three conditions where bead amount was varied for post-ligation and post-UNG purifications involve a distinct UNG step. The fourth condition also has a separate UNG step but the reaction is used as a template for PCR without purification in between. The next condition combines UNG and PCR reactions where as the last condition omits the UNG treatment all together. The sizes of these libraries were calculated from fragment smear analysis using Agilent’s software. Input was 1μg UHR total RNA. c Yield comparison of libraries made using various formats of the UNG step. As in (b) but endpoint data is concentration of the final libraries. n = 3; error bars = Standard Deviation