GSM9258258: smRNAseq_Flower_b11_pRIM22_RIM22B3mut-3xFLAG_line2; Arabidopsis thaliana; ncRNA-Seq
Total RNA extraction and smRNA enrichment was performed following a previously reported protocol. Briefly, 10 μg of total RNA extracted from flower buds (stage 12 and younger), ovules (stage 12 unfertilized ovules), and anthers (stage 9) were used for each smRNA library prep. The smRNA-seq libraries were constructed using the NEBNext Small RNA Library Prep Kit (NEB, Cat# E7330L). The smRNA libraries were further purified using an 8% acrylamide gel and size selected (130-160 bp) relative to the O'RangeRuler 10 bp DNA Ladder (Thermo Scientific, Cat# SM1313). The size selected smRNA libraries were pooled for sequencing with single-end 50 bp kit (SE50) on Illumina Nextseq 2000 or Novaseq X plus platforms.