Astragalus membranaceus small RNA sequencing: seedling leaf
Total RNA was extracted using Trizol reagent (Invitrogen, CA, USA) from leaves of Astragalus membranaceus seedlings following the manufacturers instructions. The quantity and purity of total RNA were checked by using Bioanalyzer 2100 and RNA 6000 Nano LabChip Kit (Agilent, CA, USA) with RIN value >7.0. Approximately 1 g of total RNA pooled from equal amount of RNA samples from leaves and roots were used to prepare small RNA library according to protocol of TruSeq Small RNA Sample Prep Kits (Illumina, San Diego, USA). The single-end sequencing (36 bp) was performed on an Illumina Hiseq2000 at the LC Sciences (Hangzhou, China) following the manufacturers protocol.