2.11 RT-qPCR
Total RNA was extracted from the cerebral cortex on the injured side
using RNA extraction reagents (Thermo Fisher, cat. no. 12183016). The
concentration and purity of the RNA were measured, and the cDNA
synthesized by reverse transcription was amplified using a fast reverse
transcription kit ((Beyotime Institute of Biotechnology). Real-time
fluorescence quantitative PCR was used for the reaction (Bio‑Rad
Laboratories, Inc.). The samples were amplified using the following
thermocycling conditions: Initial denaturation at 95˚C for 30 sec;
followed by 40 cycles of denaturation at 95˚C for 10 sec, annealing at
60˚C for 20 sec and elongation at 95˚C for 15 sec; and a final extension
at 60˚C for 60 sec. The relative mRNA expression levels of the target
genes were calculated as the fold change of the control using the
2‑ΔΔCq method (Livak et al., 2001). Primer sequences
were used in Table 1.
Table 1: Primers sequence