GSM6718792: Leaves of uncut explants after 4 days of cutting; Acorus calamus; RNA-Seq
Total RNA from each tissue was isolated using PureLink RNA mini kit (Invitrogen) following the manufacturer's instructions. Oligo dT selection or rRNA depletion was performed on the isolated total RNA. RNA was fragmented and reverse transcription to double-strand cDNA (dscDNA) by N6 random primer. The synthesized cDNA was subjected to end-repair and then was 3′ adenylated. Adaptors were ligated to the ends of these 3′ adenylated cDNA fragments. The ligation products were purified and PCR amplification was performed to enrich the purified cDNA template using PCR primer. The PCR product was denatured by heat and the single-strand DNA is cyclized by splint oligo and DNA ligase.