According to the manufacturer’s protocol, total RNA was extracted with TRIzol Reagent (Invitrogen, USA), and then was reverse-transcribed with HiScript II Q RT SuperMix for qRT-PCR (Vazyme, China) (3 (link)). An iQTM 5 Optical Module Real-Time PCR Detection System (Bio-Rad, USA) was used to perform qRT-PCR. According to the manufacturer’s instructions, the ChamQTM SYBR qPCR master mix (Vazyme biotech, China) was used to quantify mRNA. The primers used are listed in Table S1
Iqtm 5 optical module real time pcr detection system
Manufactured by Bio-Rad
Sourced in United States
The IQ™ 5 Optical Module Real-Time PCR Detection System is a laboratory instrument designed for real-time polymerase chain reaction (PCR) analysis. The core function of this system is to detect and measure fluorescent signals generated during the PCR process, allowing for quantitative analysis of nucleic acid samples.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (6 mentions)
Chamq sybr qpcr master mix, by Vazyme (3 mentions)
Hiscript 2 q rt supermix, by Vazyme (2 mentions)
Hiscript 2 q rt supermix for qrt pcr gdna wiper, by Vazyme (2 mentions)
Hiscript 2 q rt supermix for qrt pcr, by Vazyme (1 mentions)
Sybr premix ex taq 2 kit, by Takara Bio (1 mentions)
6 protocols using iqtm 5 optical module real time pcr detection system
1
Quantification of mRNA Expression
2
Total RNA Extraction and qRT-PCR Analysis
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Total RNA was extracted with TRIzol Reagent (Invitrogen, 15596026, Carlsbad, California, USA) according to the manufacturer's protocol [24 (link)]. Total RNA (1 mg) was reverse-transcribed with HiScript II Q RT SuperMix for quantitative real-time PCR (qRT-PCR) (R223-01, Vazyme, Nanjing, China). An iQTM 5 Optical Module Real-Time PCR Detection System (Bio-Rad, USA) was used to conduct qRT-PCR. ChamQTM SYBR qPCR Master Mix (Q311-02, Vazyme Biotech, Nanjing, China) was used to quantify mRNA according to the manufacturer's instructions. The primers used are listed in Supplementary Table 1 . The mRNA levels were normalized to β-actin and calculated using 2 − ΔΔCt method.
3
Quantitative RT-PCR Gene Expression Analysis
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Extracted Total RNA with TRIzol Reagent (Invitrogen) in accordance to the manufacturer's instructions. Hiscript II Q RT SuperMix for qRT‐PCR (+gDNA wiper) (Vazyme) was used to reverse transcribe the RNA (1 mg). qRT‐PCR was performed with the iQTM 5 Optical Module Real‐Time PCR Detection System (Bio‐Rad). mRNA was quantified using the ChamQtm SYBR qPCR master mix according to the manufacturer's instructions. The 2 - Δ Δ C t
4
Quantitative Gene Expression Analysis
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Total RNA was extracted from tissues with TRIzol reagent (Invitrogen, USA) following the manufacturer's protocol. Total RNA (1 mg) was reverse transcribed to generate cDNA with HiScript II Q RT SuperMix for RT‐qPCR (+gDNA Wiper) (Takara, Japan). The primer sequences are listed in Table S1 (Supporting Information). RT‐qPCR was performed on the iQTM 5 Optical Module Real‐Time PCR Detection System (Bio‐Rad, China) using a SYBR Premix Ex Taq II kit (Takara, Japan). The 2‐ΔΔCT method was used to quantitate changes in relative gene expression normalized to β‐actin.
5
qRT-PCR Analysis of Gene Expression
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qRT-PCR was performed as previously described [27 ]. Total RNA was isolated from cells using TRIzol reagent (Invitrogen). Reverse transcription was conducted to obtain cDNAs using HiScript II Q RT SuperMix for qRT-PCR (+gDNA wiper) (Vazyme, USA). qRT-PCR was conducted using an iQTM 5 Optical Module Real-Time PCR Detection System (Bio-Rad, USA). Gene expression was normalized to β-actin and calculated using the 2-ΔΔCt method. The primer sequences are listed in Table S1.
6
Total RNA Extraction and qRT-PCR Analysis
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Total RNA was extracted using TRIzol Reagent (Invitrogen, Carlsbad, California, USA) in accordance with the manufacturer’s protocol. Subsequently, 1 mg of total RNA was reverse-transcribed using the HiScript II Q RT SuperMix for quantitative real-time PCR (qRT-PCR) (R223-01, Vazyme biotech). The qRT-PCR was performed using an iQTM 5 Optical Module Real-Time PCR Detection System (Bio-Rad, USA). ChamQTM SYBR qPCR master mix (Q311-02, Vazyme biotech) was utilized to quantify mRNA as per the manufacturer’s instructions. The mRNA levels were normalized to β-actin and calculated using the 2-ΔΔCt method.
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