GSM8796553: sRNA_clsy12ov1; Arabidopsis thaliana; ncRNA-Seq
Pistils and anther (stage 9 or younger) of each genotype were collected and ground into fine powder with liquid nitrogen. Total RNA was extracted using the Direct-zol RNA Miniprep kit (Zymo) according to the manufacturer's instructions. Two micrograms of total RNA were mixed with equal volume of the 2x RNA loading dye, denatured at 65◦C for 10 min and immediately chilled on ice. Denatured total RNA was separated on 15% TBE urea gel (Invitrogen) and small RNAs from 15-30 nts were excised. The excised gel pieces were mashed by pestles. Small RNA was eluted with 400 uL nuclease-free water at 70◦C for 10 mins and then precipitated with ethanol. Small RNA libraries were made using NEBNext® Small RNA Library Prep Set for Illumina® (Multiplex Compatible) according to the manufacturer's instructions