title: GSM4705970: Innercork April replicate 3 [ap3_11164_CAGATC]; Quercus suber; RNA-Seq
library_construction_protocol: Total RNA was extracted from cork tissue by a modification of the method described by Chang et al. (1993). Five g of tissue was ground in liquid nitrogen and then mixed with extraction buffer (2% CTAB, 2% PVP, 10 mM Tris-HCl pH 8.0, 25 mM EDTA, 2 M NaCl and 2.67% 2-mercaptoethanol). After a 10-min incubation at 65 °C, two chloroform:isoamyl alcohol 24:1 (v/v) extractions at room temperature were performed, followed by an overnight precipitation with 2 M LiCl at 4 °C. The precipitate was collected by centrifugation, and the dry pellet was resuspended in buffer containing 1 M NaCl, 0.5% SDS, 10 mM Tris-HCl pH 8.0 and 1 mM EDTA and incubated at 60 °C for 5 min. Then, one acid phenol:chloroform:isoamyl alcohol (125:24:1 (v/v)) and two 24:1 (v/v) chloroform:isoamyl alcohol extractions were performed at room temperature. The RNA in the upper phase was precipitated with 2 volumes of 100% ethanol and the precipitate collected by centrifugation. The pellet was washed twice with 70% (v/v) ethanol and resuspended in 50 µl of RNase-free water. RNeasy MinElute Cleanup (Qiagen) was used to remove any remaining protein fragments and genomic DNA was on-colum digested with DNase I (Qiagen). The cDNA libraries were prepared using the TruSeq RNA Library Prep Kit (Illumina)
attributes.tag: GEO Accession
attributes.value: GSM4705970