design: cDNA library prepared via TruSeqTM RNA Sample Preparation Kit (Illumina): 1g total RNA  oligo(dT) beads isolate mRNA  fragmentation buffer fragment mRNA  cDNA synthesis  end repair + A-base addition  Illumina-indexed adaptors ligation  2% agarose size selection (200-300bp)  Phusion DNA polymerase PCR (15 cycles)