title: GSM7866277: Garlic-1_S12_L001; Allium sativum; miRNA-Seq
library_construction_protocol: ENVs were isolated from soy bean, ginger, amla and turmeric using a cost effective Polyethylene glycol- based (PEG) precipitation method developed in our lab (Kalarikkal et al., Scientific Reports, 2020). In brief, edible portion of plants were ground in phosphate buffered saline (PBS) using a kitchen mixer. In the case of Soybeans, they were soaked in PBS for overnight prior to grinding. Aqueous extracts were subjected to three rounds of centrifugation at 1200g for 20 min (remove cells/nuclei), 3000g for 20 min and 10,000g for 45 min (remove fibers and insoluble contents). This is known as S10 extract (crude extracellular extract). To purify ENVs, S10 extract was subjected precipitation with 10% PEG-6000, overnight followed by centrifugation at 10000g for 30 min. ENV pellets are suspended in sterile water and stored at -80 in aliqouts. ENVs were quantified by measuring the lipids by vanillin assay. Total RNA was isolated from two independent batches of ENVs (5mg lipid equivalent) for each plant by TRI RNA extraction reagent (Sigma Aldrich) as per manufacturers protocol. 1 μg of total RNA was subjected to library preparation