small RNA-Seq of Arabidopsis thaliana: Col-0 whole seed, germination stimulated
Small RNA-Seq libraries were constructed following in-house protocols (Shin et al., 2018). Briefly, 18- to 30-nt small RNAs were selected from 15 ug of total RNA of each samples by 15% urea-PAGE. Eluted size-selected small RNAs were ligated into 3-adapter and 5-adapter sequentially, followed by urea-PAGE separation for each step of adapter ligation reaction. Adapter-ligated small RNAs were amplified using Phusion Polymerase (NEB) and then 130~150-bp of amplified products were separated by native-PAGE. Eluted small RNAs were analyzed using HiSeq2500 with 51-cycle single-end mode by Macrogen (Seoul, South Korea).