title: Illumina HiSeq 2500 paired end sequencing; RNA-seq of WT and dja6 dja5 mutant in Arabidopsis thaliana
design: RNA-seq of WT and dja6 dja5 mutant in Arabidopsis thaliana
library_construction_protocol: The total tissues from 3-week-old WT and dja6 dja5 seedlings were collected and frozen quickly in liquid nitrogen before the extraction of total RNA. Arabidopsis thaliana Columbia-0 wild type and dja6 dja5 mutant seeds were pre-incubated in darkness for 48 h at 4℃ and sowed on the Murashige and Skoog (MS) medium containing 2% sucrose after surface-sterilization. Seedlings were grown on the MS medium for 3 weeks under a photoperiod of 12 h light/12 h dark cycle with a photon flux density of 120 μmol m-2 s-1 at 22℃.  Total RNA was extracted using an RNeasy Plant Mini Kit (Cat No. 74904, Qiagen, Germany). The RNA sample was measured using Thermo NanoDrop 2000 (USA).  The RNA sample was enriched using the Oligo (dT) magnetic beads, and subsequently fragmented in fragmentation buffer and revised into cDNA library.  The samples were further purified using AMPure XP beads(Beckman Coulter, Brea CA, USA). Repair the ends of the double-stranded cDNA, tail the PCR products with adenosine and add the adapters. The sample were re-purified using AMPure XP beads. PCR amplify the cDNA.