GSM5237594: zmpnrpd1-br2 (B); Arabidopsis thaliana; miRNA-Seq
Inflorescences were collected and frozen in liquid nitrogen and kept at -80oC unitl use.  Total RNA was extracted using TRI reagent (MRC) according to the manufacturer's protocol, and resolved in 15% urea-polyacrylamide gels, from which gel pieces corresponding to the 10-50 nt small RNA fraction were excised. The small RNAs were recovered by soaking the gel slices in 0.4 N NaCl followed by ethanol precipitation. The resulting small RNAs were then used for library preparation. Extracted small RNAs were used for library preparation with the NEBNext Multiplex Small RNA Library Prep Set for Illumina (New England Biolabs) following the manufacturer's protocol. The final library products were resolved on a 12% polyacrylamide gel, from which the 150 bp band as determined by the pBR322 DNA-MspI Digest ladder (New England Biolabs, E7323AA, provided in NEB 7300) was excised. The libraries were pooled and sequenced.