title: GSM2151937: RNA WT low Pi rep3; Arabidopsis thaliana; RNA-Seq
library_construction_protocol: Genomic DNA was isolated from rosette leaves using a Wizard Genomic DNA Purification kit A1120 (Promega, USA). The procedure involves lysis of isolated nuclei, RNase digestion, protein precipitation, centrifugation, precipitation of DNA from the supernatant using isopropanol, followed by a wash step with 70% ethanol. After pelleting the precipitated DNA by centrifugation, it was dried under a vacuum and rehydrated in sterile nuclease free water. Total RNA was isolated from rosette leaves using a Plant total RNA Purification kit Cat#TR02 (Gene Mark, Taiwan). The procedure involves cell lysis and protein precipitation followed by centrifugation. RNA in the supernatant is then bound on a filter column, where it is digested with DNaseI, followed by washing, and eluting with nuclease-free waster. BS-seq: Genomic DNA was sheared by sonication to generate DNA fragments in the 100-300 bp size range. Bisulfite treatment and library preparation were carried out as described (Feng et al. 2011), except that the invitrogen methylcode bisulfite conversion kit was utilized for bisulfite treatment. The resulting libraries were sequenced with  Illumina sequencing technology. RNA-seq: RNA was quantified with the Qubit RNA BR Assay and 10 μg of purified RNA was utilized as starting material for library construction, which was carried out with the Dynabeads for polyA RNA enrichment, Superscript III cDNA synthesis, and barcoded with illumina index.  Libraries were sequenced using 100-bp reads on the Illumina System.
attributes.tag: GEO Accession
attributes.value: GSM2151937