Total RNA was extracted using Column Animal RNAout kit (Tiandz) according to the manufacturer’s instructions. PrimeScript RT reagent Kit with gDNA Eraser (Takara) was used for reverse transcription-PCR reaction. Gene expression was quantified by QuantiNova SYBR Green PCR Kit (Qiagen) using the ABI Prim 7300 Sequence Detection System (Applied Biosystems). The relative expression of mRNA was calculated by normalization to GAPDH relative to the control.
Abi prim 7300 sequence detection system
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The ABI Prism 7300 Sequence Detection System is a real-time PCR instrument designed for quantitative gene expression analysis. It utilizes fluorescence detection technology to monitor the amplification of DNA sequences during the PCR process. The system provides accurate quantification of target nucleic acids and supports a wide range of applications, including gene expression profiling, SNP genotyping, and viral load determination.
Automatically generated - may contain errors
2 protocols using abi prim 7300 sequence detection system
1
RNA Extraction and qRT-PCR Protocol
2
Quantitative mRNA Analysis by RT-PCR
Check if the same lab product or an alternative is used in the 5 most similar protocols
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For analysis of mRNA, HiScript 1st strand cDNA Synthesis Kit (Vazyme Biotech Co., Nanjing, China) was used for RT-PCR reaction. Gene expression was quantified by SYBR Green Q-PCR Kit (Roche, Germany) using the ABI Prim 7300 Sequence Detection System (Applied Biosystems, Foster city, CA). Specific primers for mRNAs are listed in Supplementary Table 1 . The Ct values were analyzed using the ΔΔCt method and relative changes of mRNA levels were obtained by normalization to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) relative to the control.
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