Small RNA sequencing of Andrographis paniculata
To extract the RNA, tissues were collected from the RNAlater solution and submerged into the Liquid Nitrogen. Later, the frozen tissues were manually grind to fine powdered form by using mortal and pestel. The finely grinded samples were weighed to 500 mg for RNA extraction. One mL of the CTAB buffer (Hexadecyltrimethylammonium bromide 2% W/V; PVP-40 2% W/V; 100 mM TrisHCl pH 8.0; 25 mM EDTA, 2 M NaCl; and 2% -mercaptoethanol fresh) was used for further lysis of the cells at 65  for 30 min. Further for RNA precipitation overnight at 4 , 1/10th volume of lithium chloride (LiCl) was added following the addition of equal amount of chloroform:isoamylalcohol (24:1) over supernatant and supernatant was collected after 5 min of incubation. Eventually, the precipitated RNA was washed with 70% ethanol (EtOH) and RNA was resuspended in 30 uls of RNase free Water. It was further subjected to DNA removal by using Turbo DNAse by following manufacturers protocol. Consequently, to assure the quality of extracted RNA, Nanodrop 1000 spectrometer was used and for RIN value calculations Agilent Tapestation was utilised.