Quantitative PCR Analysis of RNA Expression

After the total RNA was extracted from BV2 cells, primary microglial cells or tissues using RNAiso Plus reagent (Takara, Tokyo, Japan), the total RNA (500 ng) was reverse transcribed into cDNA using HiScript III first Strand cDNA Synthesis Kit (Vazyme, Nanjing, China). Real-Time Quantitative PCR was carried out using Taq Pro universal SYBR qPCR Master Mix (Vazyme, Nanjing, China) and ABI-7500 quantitative PCR system (Applied Biosystems, Warrington, United Kingdom). The sequence of PCR primers was shown in Table 2 (Gu et al., 2018 (link)).

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Fang J., She J., Lin F., Wu J.C., Han R., Sheng R., Wang G, & Qin Z.H. (2022). RRx-001 Exerts Neuroprotection Against LPS-Induced Microglia Activation and Neuroinflammation Through Disturbing the TLR4 Pathway. Frontiers in Pharmacology, 13, 889383.

Publication 2022

RNAiso Plus reagent HiScript III first Strand cDNA Synthesis Kit Taq Pro universal SYBR qPCR Master Mix ABI-7500 quantitative PCR system

Cdna Microglial cells Primers Real time pcr Synthesis Tissues

Corresponding Organization :

Other organizations : Beijing National Laboratory for Molecular Sciences

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Gene expression measured by Real-Time Quantitative PCR

control variables

RNA extraction using RNAiso Plus reagent
Reverse transcription into cDNA using HiScript III first Strand cDNA Synthesis Kit
PCR amplification using Taq Pro universal SYBR qPCR Master Mix
PCR system: ABI-7500 quantitative PCR system

controls

Positive control: Not specified
Negative control: Not specified

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