GSM8389280: ADF8_RNA_0h_Pi_rep3; Arabidopsis thaliana; RNA-Seq
Biotin tagged nuclei from ~100 mg from root tissue 1 cm above the root tip were captured on 25 uL of Streptavidin M280 Dynabeads according to the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol . Bead bound nuclei for each timepoint in the starvation assay were counted in 1 mL of Nuclei Purification Buffer with 0.1% Triton X-100 (NPBt) and nuclei were split three ways to ~100,000 nuclei per sample in NPBt for three replicates. RNA was isolated from bead bound nuclei using the RNeasy plant micro kit (Qiagen) with on column DNase treatment and quantified via Quant-it™ RiboGreen RNA Assay Kit (Thermo-Fisher). Libraries for RNA-seq were generated according to the QuantSeq 3' mRNA-Seq Library Prep Kit FWD V1 (Lexogen, cat.no. 015.24) following modified steps for low input samples.