GSM9027442: Altenb-2, control, rep2; Arabidopsis thaliana; RNA-Seq
Total RNA was extracted using Sepasol-RNA I Super G (Nacalai Tesque) following the manufacturer's protocol. Then diluted RNA was subjected to Phenol/Chloroform precipitation, LiCl precipitation, and isopropanol precipitation.The concentrations of the extracted RNA samples were measured with a spectrophotometer (BioSpectrometer Basic; Eppendorf), and 10 ng of total RNA was used to CAGE-seq. The cDNAs were synthesized from total RNA using random primers. The ribose diols in the 5' cap structures of RNAs were oxidized, and then biotinylated. The biotinylated RNA/cDNAs were selected by streptavidin beads (cap-trapping). After RNA digestion by RNaseONE/H and adaptor ligation to both ends of cDNA, double-stranded cDNA libraries (CAGE libraries) were constructed.