Generate a highly representative cDNA pool for sequencing on Illumina instruments.
Electropherogram of amplified SMARTer cDNA. Various amounts of Universal Human Reference Total RNA (UHR) and Human Brain Reference RNA were used as input for SMARTer cDNA synthesis.The cDNA samples were then analyzed for purity and yield on an Agilent 2100 Bioanalyzer. Shown are Bioanalyzer trace overlays of cDNA amplified from 1 ng (red line), 0.1 ng (dark blue line), 0.05 ng (green line), and 0.01 ng (light blue line) of total RNA and a no template control (NTC; pink line). The main peak indicates the purity and yield of cDNA between 0.4 and 9 kb—with the highest point at ~2 kb. There was no amplification in the negative control (pink line). Although the amount of input RNA can vary over quite a large range (e.g., 1 ng to 0.01 ng), comparable cDNA output can be obtained by adjusting the number of PCR cycles.