tier27_row3_tree46_Jun12_apices_rep1
RNA from summer apices was extracted using a previously-developed protocol treated with a DNA-freeTM Kit (Thermo Fisher Scientific, Waltham, MA) to remove DNA, and assessed for initial quality using a Nanodrop OneC (Thermo Fisher Scientific, Waltham, MA) (Gasic et al., 2004). If purity was insufficient (260/230 < 1.8), samples were further cleaned and concentrated using the RNA Clean & ConcentratorTM-25 kit (Zymo Research, Seattle, WA). Concentration of each sample was measured using the Qubit RNA Broad Range kit and Qubit 4 Fluorometer (Thermo Fisher Scientific Inc, Waltham, MA), and quality was assessed with an Agilent High Sensitivity RNA ScreenTape System (Agilent Technologies, Santa Clara, CA). Illumina stranded mRNA 150 bp PE libraries were prepared at MSUs RTSF before they were pooled and sequenced on approximately 0.8 lanes of a NovaSeq 6000, producing at least 39.8 million reads per library.