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.