Small RNA-seq libraries were prepared from total RNA extracted using TRIzol  from 100 mg of root tissue. RNA was resuspended in 10 µl of 10 mM Tris (pH 7.0) and separated on a 7 M urea-denaturing  polyacrylamide gel electrophoresis (PAGE) in a 15% polyacrylamide gel. Marker oligos were used to demarcate fragments of 18-28 nt, which were collected. Gel slices were processed in 250 µl gel extraction buffer [20 mM Tris (pH 7.0), 250 mM NaOAc, 1 mM; EDTA, 0.25% (w/v); SDS] by freezing at -80°C for at least 30 min and then incubation for 15 min at 23-25 °C with rocking. The eluate was recovered by centrifugation for 10 min at 13,000 x g and the RFs recovered by ethanol precipitation. sRNA libraries were prepared using NEXTflex™ Small RNA Sequencing Kit (BIOO Scientific).
GEO Accession
GSM2601712