2.8 RNA quality assessment, Illumina sequencing, and DEGs analysis
Advanced molecular biology techniques were applied to assess the concentration and quality of the RNA sample. The RNA that met the qualifications was t used for library construction. To ensure library quality, the cDNA concentration and insert size were assessed using Qubit 2.0 and Agilent 2100. The effective concentration of the library (>2 nM) was accurately measured through qPCR. Upon confirmation of the library’s suitability, the pooling and sequencing were performed using the Illumina platform. The selection criteria for DEGs were established as FC ≥ 2 and FDR < 0.01(Anders and Huber, 2010). The FDR refers to the adjusted p-value used to determine the significance of the difference.