qRT-PCR was used to determine the mRNA level of HIF-1α, PDK1, GLUT1, HK2, and LDHA in cerebral cortex. TRIzol reagent (Vazyme, Nanjing, China) was used to extract total RNA from the cortical tissue following the manufacturer's instruction. The purity and concentration of RNA were determined by ultraviolet spectrophotometry. Reverse transcription of RNA was performed using the Reverse Transcriptase kit (Bimake, Houston, TX, USA). QRT-PCR was conducted to analyze the gene expression using the primer (Sangon Biotech, Shanghai, China) and SYBR Green II (Bimake, Houston, TX, USA). All data were normalized by β-actin. The primer sequences are shown in Table 1 .
Sybr green 2
Manufactured by Selleck Chemicals
Sourced in China, United States
SYBR Green II is a nucleic acid stain used in molecular biology applications. It is a fluorescent dye that binds to double-stranded DNA, allowing for the detection and quantification of DNA in various assays, such as real-time PCR and gel electrophoresis.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sybr green 2
1
Quantifying Cortical Hypoxia-Responsive Genes
2
Quantitative Transcription Analysis
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Total RNA was isolated from cells in the control group, GF group, and LXR+GF group by Trizol reagent (Vazyme, Nanjing, China) according to the manufacturer’s protocol. mRNA was subjected to reverse transcription using HiScript Q Select RT SuperMix (Vazyme). SYBR Green II (Bimake, Houston, TX, USA) incorporation method was used to detect the amount of mRNA. Negative controls were used as no template cDNA reactions and melting curves were used to confirm the results. The results were normalized using the glyceraldehyde 3-phosphate dehydrogenase (GAPDH) concentration of each sample. The primer sequences are reported in Table 1 .
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