GSM8464430: hmr-5 27℃ w/o suc rep2; Arabidopsis thaliana; RNA-Seq
100 mg seedlings were collected by flash freezing in liquid nitrogen at the indicated time point. Plant tissues were ground to a fine powder in liquid nitrogen, and total RNA was extracted using Quick-RNA MiniPrep kit with on-column DNase I digestion (Zymo Research). Library construction was performed by Novogene. Ribosomal RNA was removed from total RNA, followed by ethanol precipitation. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. During the second strand cDNA synthesis, dUTPs were replaced with dTTPs in the reaction buffer. The directional library was ready after end repair, A-tailing, adapter ligation, size selection, enzyme digestion to remove UTP-containing second strand cDNA, amplification, and purification. The libraries were subjected to paired-end 150 bp sequencing using NovaSeq 6000