GSM3620632: polysome NAD_P rep1; Arabidopsis thaliana; OTHER
Total RNAs were extracted from 12-day-old seedlings and inflorescences by the Trizol reagent, and then treated with DNase I (Roche; 1U /μgRNA at 37 °C for 60 min). NEBNext® Ultra™ Directional RNA Library Prep Kit was used for subsequent RNA library preparation. The First Strand Synthesis Reaction Buffer (containing zinc ions to fragment RNA) was mixed with the random primers to make the reverse transcription mixture (RT mixture). The beads with bound NAD+-capped RNAs were suspended with the RT mixture and incubated at 94°C for 6 min to 1) elute the RNAs from beads, 2) fragment the RNAs, and 3) denature the RNAs. Then RNase Inhibitor and reverse transcriptase were added to initiate first strand cDNA synthesis. The remaining steps were performed according to manufacturer's instructions NAD+- capture RNA seq
GEO Accession
GSM3620632