RNA-Seq of  Arabidopsis thaliana: pollen
The  total RNA samples are first treated with DNase I. Then the mRNA is enriched  by using the oligo(dT) magnetic beads. Mixed with the fragmentation buffer,  the mRNA is fragmented into short fragments. Then the first strand of cDNA  is    synthesized by using random hexamer-primer. Buffer, dNTPs, RNase H and DNA  polymerase I are added to synthesize the second strand. The double strand  cDNA is purified with magnetic beads. End    reparation and 3'-end single nucleotide A (adenine) addition is then performed.  Finally, sequencing adaptors are ligated to the fragments. The fragments are  enriched by PCR amplification. The library products are ready for sequencing  via Illumina    HiSeq 2000