GSM8796107: sRNA_Nodule_3wpi_2; Glycine max; miRNA-Seq
Frozen nodules were manually ground using pre-chilled (liquid N2) mortar and pestles. Ground tissue was further pulverized using a MM400 tissuelyzer (Retsch, Germany): 2x 45 sec at 30 Hz. Total nodule RNA was extracted using TRIreagent (Ambion, USA), following manufacturer's instructions plus additional isopropanol rinse to remove the remaining polysaccharides (Protocol 1: Soybean Nodule RNA Extraction). sRNA libraries were constructed using the RealSeq-AC kit version 2 (Realseq Biosciences, Santa Cruz, CA, catalog no. 500-00048;) as per the manufacturer's instructions. For RNAseq libraries, we used NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs, catalog no. E7760L) protocol.