title: RNA-Seq of CD8+ Tcell from spleen of mice
design: Enrichment of eukaryotic mRNA using magnetic beads with Oligo(dT). Then fragmentation buffer is added to break the mRNA into short fragments. Using the mRNA as a template, six-base random primers (random hexamers) are used to synthesize one-strand cDNA. Then buffer, dNTPs, and DNA polymerase I and RNase H are added to synthesize two-strand cDNA. Double-stranded cDNA was then purified using AMPure XP beads. Purified double-stranded cDNA was end-repaired, A-tailed, and sequencing adapters ligated, followed by fragment size selection using AMPure XP beads. Finally, PCR amplification was performed, and the PCR product was purified with AMPure XP beads to obtain the final library.