mac7_CLCuDfree_6
Poly (A) mRNA was isolated using oligodT beads (Qiagen, Hilden, Germany). The mRNA was broken into short fragments (~300 nt). Firststrand cDNA was synthesized using random hexamerprimed reverse transcription. Secondstrand cDNA was generated using RNase H and DNA polymerase I. The cDNA fragments were purified and washed for end repair and ligated to sequencing adapters. The cDNA fragments of suitable size were purified and enriched by PCR to obtain the final cDNA library. The integrity of each cDNA library was examined using Bioanalyzer 2100 equipment (Agilent Technologies, Germany).