NextSeq 500 paired end sequencing; Meiotic cohesin, chromatin and expression profiling of Arabidopsis thaliana floral buds from wild type and non-CG DNA methylation/H3K9me2 pathway mutant plants (RNA-seq)
Meiotic cohesin, chromatin and expression profiling of Arabidopsis thaliana floral buds from wild type and non-CG DNA methylation/H3K9me2 pathway mutant plants (RNA-seq)
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. 6 μg of total RNA was treated with DNAseI using TURBO DNase reagents (ThermoFisher, AM2238). 4 μg of total DNAseI-treated total RNA was then treated with Ribo-Zero rRNA removal magnetic kit (Epicentre, MRZPL116) to deplete rRNA from the total RNA sample. 100 mg of unopened flower buds were ground in liquid nitrogen and RNA was extracted using Trizol reagent. 50 ng of rRNA-depleted DNAseI-treated RNA was used to prepare a stranded RNA-seq library using the ScriptSeq v2 RNA-seq Library Preparation Kit (Epicentre, SSV21124), as described in Choi et al. (2018) Genome Res. 28(4): 532-546.
Experimental Factor: genotype
wild type genotype