GSM7728588: Spleen_CK_rep2_RNA; Gallus gallus; RNA-Seq
Total RNA of the spleen in each group was extracted by using TRIzol® RNA extraction reagent (Invitrogen, USA) according to the manufacturer's instructions. The integrity of RNA was examined by agarose gel electrophoresis, and the concentration of RNA was measured by NanoDrop 2000 spectrophotometers (Thermo Fisher, USA). The mRNA was purified by oligo (dT) magnetic beads and fragmented into short fragments using fragmentation buffer. First-strand cDNA was synthesized with SuperScript II Reverse Transcriptase (Invitrogen) using random primers, and second-strand cDNA was synthesized using the synthesized first strand of cDNA as a template. The obtained double-stranded cDNA was purified by a VAHTS® mRNA-seq V3 Library Prep Kit for Illumina (Vazyme, China), end repaired, poly(A) added, and then ligated to Illumina sequencing adapters. Sequencing was performed on the Illumina HiSeq-2000 platform for 50 cycles. High-quality reads passed through the Illumina quality filter were retained in fastq.gz format for sequence analysis.