GSM9060195: biol rep2 M_3H_A; Arabidopsis thaliana; RNA-Seq
Arabidopsis tissue samples were subjected to total RNA extraction via the RNeasy Plant Mini Kit (QIAGEN), incorporating an on-column DNase I step for genomic DNA removal. The RNA-Seq library construction and sequencing services were provided by Novogene Co., Ltd. (Beijing, China), with three biological replicates per sampleA total RNA preparation was cleaned of ribosomal RNA, followed by ethanol precipitation. After fragmentation, the first strand cDNA was synthesized using random hexamer primers. During 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, USER enzyme digestion, amplification, and purification. The library was checked with Qubit and the subjected to real-time PCR for quantification and a bioanalyzer for detection if fragment size distribution. Quantified libraries were pooled and sequenced on Illumina NovaSeq 6000 platforms using a PE150 read length, with over 12 G of raw data per sample were generated.