GSM1528218: S-7d; Salicornia europaea; miRNA-Seq
Plant tissues were ground in liquid nitrogen and were added into two volume of extraction buffer containing 2 M NaCl, 25 mM EDTA, 200 mM Tris (pH 8.0), 20 mM sodium borate, 2% PVPP (w/v), 2% CTAB (w/v), 1% LSS (w/v), and 2% b-mercaptoethanol (v/v). The homogenate was then transferred into a sterile disposable polypropylene tube, vortexed vigorously, and incubated at 65°C in a water-bath shaker over 10 min with occasional shaking. Added one volume of chloroform/isoamyl alcohol (24:1, v/v) into the mixture, and then vortexed, followed by centrifuging at 15,000g for 15 min at 4°C. The resulting upper aqueous phase was transferred into another tube containing one volume of water-saturated phenol/chloroform/ isoamyl alcohol (25:24:1, v/v, pH 4.5). Following vortexing, the sample was then centrifuged at 12,000g for 10 min at 4°C. The above step was repeated until a clean interface was observed. The upper phase containing RNA extract was extracted with one volume of chloroform/isoamyl alcohol (24:1, v/v) and centrifuged at 12,000g for 10 min at 4°C. After the upper phase was transferred into another centrifuge tube, a volume of isopropyl alcohol was added into the tube. The mixture was incubated at-20 °C for at least 1 h or sometimes overnight, followed by centrifugation at 15,000g for 15 min at 4°C. Decant the supernatant immediately. The pellet was rinsed with 75% ethanol and collected by centrifugation at 12,000g for 5 min at 4°C for three times. The air-dried pellet was finally dissolved in DEPC-treated water and stored at -80°C. RNA libraries were prepared for sequencing using standard Illumina protocols
GEO Accession
GSM1528218