Oligonucleotide pcr primers

Yeast strains used in this study are listed in Table 1. Standard culture media and genetic techniques were used expect where noted. Escherichia coli strains DH12S (Life Technologies, Gaithersburg, MD) and DH5α (TaKaRa, Japan) were used as hosts for plasmid manipulation. SRG-Ura medium containing 2% galactose and 1% raffinose was used to overexpress genes driven by the GAL1 promoter. SC-Ura medium contained 2% dextrose. Oligonucleotide PCR primers were purchased from Sangon Biotech (Shanghai, China).

We isolated total RNA from traumatic brain tissue using Trizol reagent (Invitrogen, USA) with a sample size of (n = 3 per group). Following this, the extracted RNA was reverse transcribed using a commercially accessible Takara kit to produce complementary DNA (cDNA). Following the manufacturer’s instructions, we conducted quantitative real-time PCR (qRT-PCR) using SYBR Green PCR Master Mix (TransGen Biotech, China) and performed the PCR on a CFX96 real-time PCR detection system. The amplification protocol consisted of an initial denaturation step at 95°C for 30 seconds, followed by 40 cycles comprising denaturation at 95°C for 10 seconds, annealing at 60°C for 30 seconds, and extension at 60°C for 30 seconds. To standardize the mRNA expression levels of the target genes, we used GAPDH as the reference gene and analyzed the data utilizing the 2^−ΔΔCT method. The oligonucleotide PCR primers were procured from Sangon Biotech (Shanghai, China), and comprehensive information can be located in Table 1.