GSM6704663: Wild leaf, Stage 3, rep 1; Hakea prostrata; RNA-Seq
Frozen ground-up powder (100 mg) was mixed with Tris-HCl, LiCl, EDTA, lithium dodecyl sulfate, dithiothreitol, polyvinylpyrrolidone, polyvinylpolypyrrolidone, followed by Fruit-mate (Takara Biotechnology). After several chloroform:isoamyl alcohol extractions, RNA was concentrated by isopropanol precipitation, followed by another precipitation of the resuspended RNA in LiCl. The RNA pellet was washed with LiCl followed by two washes with ethanol, air dried, and resuspended in RNase free water. After removal of contaminating DNA (RQ1 DNase; Promega Corporation), the three replicates with the highest quality for each leaf developmental stage were selected for library preparation and sequencing. Libraries for RNA-Seq were prepared from 1 µg RNA according to the manufacturer's instructions (TruSeq Stranded Total RNA Kit with Ribo-Zero Plant, Illumina)