Hifair 2 1st strand cdna synthesis supermix for qpcr with gdna digester plus

Equal amounts of each RNA sample were used for the qRT-PCR and transcriptome sequencing analyses. The primer pairs (Supplementary Table S1) for the candidate genes were designed using Primer Premier 6.0. The total RNA was reverse transcribed using the Hifair^® II 1st Strand cDNA Synthesis SuperMix for qPCR (with gDNA digester plus) (Yeasen Biotechnology, Shanghai, China). The qRT-PCR analysis was performed using the CFX Connect Real-Time PCR instrument (Bio-Rad, Berkeley, CA, USA) and the Hieff UNICON^® Universal Blue qPCR SYBR Green Master Mix (Yeasen Biotechnology). The PCR program was as follows: 95 °C for 60 s, 40 cycles of 95 °C for 15 s, and 60 °C for 35 s. Relative gene expression levels were calculated according to the 2^−ΔΔCt method [40 (link)].

The expression of key DEGs was analyzed by qRT-PCR. Total RNA was reverse transcribed using the Hifair® II 1st Strand cDNA Synthesis SuperMix for qPCR (with gDNA digester plus) (Yeasen Biotechnology, Shanghai, China). The primer pairs (Table S1) for the candidate genes were designed using Primer Premier 6.0. The qRT-PCR analysis was performed using the CFX Connect Real-Time PCR instrument (Bio-Rad, USA) and the Hieff UNICON® Universal Blue qPCR SYBR Green Master Mix (Yeasen Biotech-nology). The 18 S gene served as an internal reference in this study. Relative gene expression was calculated using the 2^−∆∆Ct method [13 (link)].