Transcriptome of male gametophyte at a vegetative phase
Total RNA samples are first treated with DNase I to degrade any possible DNA contamination. Then the mRNA is enriched by using the oligo(dT) magnetic beads and fragmented into short fragments (about 200 bp). Then the double strand cDNA is synthesized by using random hexamer-primer. 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 HiSeqTM 2000.