2.8 Reverse transcription q-PCR validation of DEGs
In this work, to elucidate the validity of the RNA-seq data, quantitative reverse-transcription polymerase chain reaction (q-RT-PCR) was performed for some of the DEGs. The information of primers is presented in Table 4. Total RNA remaining from RNA-seq library preparation was utilized for q-RT-PCR. RNA was reverse transcribed into cDNA utilizing M-MLV Reverse Transcriptase (Vazyme). Real-time PCR was performed with the StepOne RealTime PCR System using the SYBR Green PCR Reagents Kit (Yeasen). The PCR conditions compose of denaturing at 95˚C for 10 min, 40 cycles of denaturing at 95˚C for 15 s, annealing and extension at 60˚C for 1 min. For each sample, PCR amplifications were performed in triplicate. The RNA expression levels of all the genes were standardized against that of GAPDH.