GSM1166175: RW; Brachypodium distachyon; RNA-Seq
Total RNA was extracted from 12-day-old ABR5 seedlings using the mirVana miRNA Isolation Kit (Ambion, Austin, USA) following the manufacturer's protocol. The mRNAs were enriched by using oligo(dT) magnetic beads and broken into short fragments (about 200 bp). The first strand cDNA was synthesized using random hexamer primer with mRNA fragments as templates. DNA polymerase I (Epicentre® Biotechnologies, Madison, USA) was used to synthesize the second strand. The double-strand cDNA was purified with the QiaQuick PCR extraction kit (Qiagen). Then, sequencing adaptors were added and the obtained fragments were purified using agarose gels and enriched by PCR amplification. The resultant products were used for sequencing analysis via Illumina HiSeq™ 2000.