SMARTer cDNA synthesis has minimal to no bias.
Comparison of transcript coverage with different amounts of input RNA. Shown are overlaid plots comparing the average read coverage from libraries made with 1 ng to 0.01 ng of mouse brain total RNA. The x-axis represents gene length normalized to 100%, where 0 is the 5’-end of each transcript and 100 is the 3’-end. The y-axis indicates the average coverage for a set of 724 genes that are moderately to highly expressed in brain tissue. The results are very consistent through the range of input RNA used, with full-length coverage of the transcripts reflecting no systematic 5’- or 3’-bias.