FIGURE 1.

Strategies for cDNA library preparation. (A) One approach involves reverse transcription with random primers first, followed by adapter ligations and sequencing (left). The other approach is to first sequentially ligate 3′ and 5′ adapters, then perform cDNA synthesis using a primer complementary to the adapter (RT-primer) followed by sequencing (right). On using RT-primer with a specific sequence, mispriming could occur due to annealing of the RT-primer to transcript sequences with some complementarity (RT mispriming). (B) Genome browser view showing enrichment of sequencing reads at a specific exon in EZH2 short RNA RIP compared to IgG IP and input controls across different cell lines and independent replicates (Rep). The inset shows sense-strand reads (red) mapping to the peak region.