title: RNA-seq of Col, h2a.w7, cmt3 and h2a.w7 cmt3 Arabidopsis thaliana floral buds
design: RNA-seq of Col, h2a.w7, cmt3 and h2a.w7 cmt3 Arabidopsis thaliana floral buds
library_construction_protocol: Unopened flower buds at floral stage 9 were collected in tubes, snap frozen in liquid nitrogen and stored at -80°C until further use. Plants were grown under long day conditions (16 hours light/8 hours dark) at 20°C. 100 mg of unopened flower buds was ground in liquid nitrogen and RNA was extracted using TRIzol reagent. Six micrograms of total RNA per sample was treated with DNase I. Four micrograms of total DNase I–treated total RNA was then treated with Ribo-Zero rRNA Removal Kit (MRZPL116, Epicentre) to deplete rRNA from the RNA sample. Fifty nanograms of rRNA-depleted DNase I–treated RNA was used to prepare RNA-seq libraries using the ScriptSeq v2 RNA-seq Library Preparation Kit (SSV21124, Epicentre)