GSM7893090: Ear, FM-S3, bulk RNA-seq, Replicate2; Zea mays; RNA-Seq
For bulk RNA-seq, RNA was extracted  using direct-zol RNA Miniprep (Zymo Research).For scRNA-seq, protoplasts were prepared as previously described (Satterlee et al., 2020). For each replicate, the freshly dissected FM-S2 or FM-S3 tissues from 15-20 ear or tassel were digested in protoplast lysis buffer for 2 hours at room temperature in the dark with gentle shaking. The mixture was then filtered twice through a 30 μm cell strainer (sysmex, 04-0042-2316) and the protoplasts were collected by horizontal centrifugation at 300 g for 3 min at room temperature. The supernatant was gently removed without disturbing the protoplast pellet. Next, the pellet was resuspended and washed two times using protoplast washing buffer (lysis buffer without enzyme) by the same centrifugation condition as before. After washing, the pellet was resuspended into 600 μl washing buffer and then filtered through the 30 μm cell strainer to remove adherent multicellular mass. Bulk RNA-seq librarys were constructed using NEBNext Ultra Directional RNA Library Prep Kit or MGIEasy RNA Library Prep Kit according to the instructions. scRNA-seq librarys were constructed using BD Rhapsody™ Whole Transcriptome Analysis Amplification Kit.