GSM2802523: p5_B_CAGE; Arabidopsis thaliana; RNA-Seq
Total RNA was extracted from A. thalinana tissues using TRIzol Reagent (Ambion) following the manufacturer’s instructions, and was treated with RQ1 DNase (Promega) to remove DNA. For each sample, 20μg of total RNA was used for CAGE-seq library preparation as described (Tsuchihara, Suzuki et al. 2009; Gowda, Nunes et al. 2010). In brief, polyadenylated mRNAs were purified and concentrated with oligo(dT)-conjugated magnetic beads (invitrogen). After treated with FastAP (Invitrogen) at 37℃ for 1h and subsequently with Tobacco Acid Pyrophosphatase (Ambion) at 37 ℃ for 1h, the decapped full length mRNA was ligated to Turseq 5' RNA adaptor (illumina) at 37 ℃ for 1h, and purified with oligo(dT)-conjugated magnetic beads (invitrogen). Following iron fragmentation at 95℃, first strand cDNA was synthesized with RT primer harboring Turseq 3' adaptor sequence (illumina) and a randomized hexamer. The cDNAs were purified and amplified with Truseq PCR primers (illumina) and PCR products corresponding to 200-500 bps were purified, quantified and stored at -80 ℃ until used for sequencing.
GEO
GSM2802523