GSM1202547: tbh_rep1; Cucumis sativus; RNA-Seq
Total RNA were extracted using Huayueyang RNA extraction kit (China) 6ug total RNA was used in library construction.Double-stranded cDNAs were synthesized using oligo(dT) beads (Invitrogen).The cDNAs were then digested with an anchoring restriction enzyme (NlaIII) and ligated to a 5' adapter, which contains a recognition site for the type IIS restriction enzyme MmeI (New England Biolabs). Following MmeI digestion, 3' adapter was ligated. Tags flanked by both adapters were enriched by PCR for 15 cycles. The PCR products were run on a 6% PAGE gel, and the 105-bp DNA band was excised and purified. The DNA samples were sequenced on an Illumina HiSeq 2000 sequencer.