title: GSM5093664: CK_R1; Gossypium hirsutum; RNA-Seq
library_construction_protocol: Total RNA was extracted using TRlzol Reagent(Life technologies, California, USA) following the manufacturer's procedure. The integrity and concentration of total RNA was checked by Agilent 2100 Bioanalyzer (Agilent Technologies, Inc., Santa Clara, CA, USA). The isolated mRNA by NEB Next Poly (A) mRNA Magnetic Isolation Module (NEB, E7490) were used for constructing cDNA library through the manufacturer's instructions of NEBNext Ultra RNA Library Prep Kit for Illumina (NEB, E7530) and NEB Next Multiplex Oligos for Illumina (NEB, E7500). Approximately 200nt RNA inserts were used to synthesize the first-strand cDNA and the second cDNA, according to the fragmented mRNA. In the next step, the end-repair / dA-tail and adaptor ligation were performed for double-stranded cDNA, which is to form the cDNA library by Agencourt AMPure XP beads (Beckman Coulter, Inc.) and PCR amplification. Finally, the constructed cDNA libraries of the different samples were sequenced on a flow cell using an Illumina Novaseq 6000 platform. The paired-end libraries was 300 bp ( ± 50 bp)