GSM8442838: vrn1 FRI, warm, bio_rep3; Arabidopsis thaliana; MNase-Seq
1.5 g of seedlings per replicate was ground with liquid nitrogen to a fine powder Nuclei were isolated and purified on a Percoll gradient. 65 mg of purified nuclei were treated with 10 U of MNase (Worthington) for 30 min at 37°C. DNA was extracted with phenol-chloroform and treated with RNase A for 20 min at 37°C. Mono- and sub-nucleosomal DNA fragments were purified with AMPure XP beads (Beckman Coulter), recovering all fragments < 200 bp and excluding all fragments > 300 bp. Sequencing libraries were prepared using the NEBNext Ultra II DNA Library Prep Kit (NEB, #E7645) following the manufacturer's instructions. After adaptor ligation, DNA was purified with AMPure XP beads, recovering all fragments > 200 bp. Illumina indexing PCR was performed with 8 cycles. Libraries were pooled and enriched for target loci in one reaction using myBaits Custom Kit (Daicel Arbor Biosciences) following the manufacturer's instructions for high sensitivity hybridization capture for targeted NGS.