GSM7775234: Leaf, Mock, 3 hours, rep 1 [preM_A1]; Arabidopsis thaliana; RNA-Seq
Total RNA was extracted using the Direct-zol RNA Miniprep Plus Kit (Zymo Research according to the manufacturer's instructions).  A total of 2ug were used for libraries preparation. RNA sequencing was performed by Novogene (Novogene Co., Ltd., Beijing, China). A total of 2000 ng total RNA was used to extract messenger RNA, using poly-T oligo-attached magnetic beads. Random hexamer primers were used to synthesize the first strand of cDNA from the fragmented RNA. The second strand was synthesized using either dUTP or dTTP, depending on whether the library was directional or non-directional. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, USER enzyme digestion, amplification, and purification. The library was quantified using Qubit and real-time PCR and detected size distribution using a bioanalyzer device. Libraries were pooled and sequenced using Illumina sequencing and paired-end reads were generated.