RNA-Seq of Ficus altissima: leaves
Total RNA was extracted using Trizol reagent kit (Invitrogen, Carlsbad, CA, USA)according to the manufacturers protocol. RNA quality was assessed on an Agilent 2100Bioanalyzer (Agilent Technologies, Palo Alto, CA, USA) and checked using RNasefree agarose gel electrophoresis. After total RNA was extracted, eukaryotic mRNA wasenriched by Oligo(dT) beads, while prokaryotic mRNA was enriched by removingrRNA by Ribo-ZeroTM Magnetic Kit (Epicentre, Madison, WI, USA). Then the enrichedmRNA was fragmented into short fragments using fragmentation buffer and reversetranscribed into cDNA with random primers. Second-strand cDNA was synthesized byDNA polymerase I, RNase H, dNTP and buffer. Then the cDNA fragments werepurified with QiaQuick PCR extraction kit(Qiagen, Venlo, The Netherlands), endrepaired, poly(A) added, and ligated to Illumina sequencing adapters. The ligationproducts were size selected by agarose gel electrophoresis, PCR amplified, andsequenced using Illumina HiSeqTM 4000 by Gene Denovo Biotechnology Co.(Guangzhou, China).
SUB9312885
FA_leaf