Total RNA from cells and tissues was extracted with TRIzol reagent (Thermo Fisher Scientific, Waltham, MA, USA). Then, 1 μg RNA was reverse transcribed into cDNA using the HiScript III 1st Strand cDNA Synthesis Kit (Vazyme Biotech, Nanjing, China). Real-time quantitative PCR was conducted using Taq Pro Universal SYBR qPCR Master Mix (Vazyme Biotech, Nanjing, China) on a CFX96 Real-Time quantitative PCR Detection System (Bio-Rad Laboratories, Hercules, CA, USA). The primer sequences were as follows: LINC01615: 5′-GAAGACAGGGGATCCCGAAG-3′ and 5′-AATGAAAGTCCAGCAGGAGGG-3′; MIR100HG: 5′-CCCAGTGCAAGGACAAAGA-3′ and 5′-GCAGAGGAGGTGTCTTCAGG-3′; LINC00702: 5′-GCAGTGGCATGATCTCGGCT-3′ and 5′-GGCCGAGGCAGGTGGATAAC-3′; LINC02544: 5′-TCTCATTCGTGGCTGGATCA-3′ and 5′-ACGCTCTCGAAATCGTGTCC-3′; SERTAD4-AS1: 5′-CCTATTCCCTGCTTCTGCGA-3′ and 5′-AGCCAGAGGTCTGGTTTTTCA-3′; FENDRR: 5′-CCACATGGATGGTTGCCACTCTC-3′ and 5′-GCTGGTACTCGGCCTTCTAATTGG-3′; GAPDH: 5′-TGAACGGGAAGCTCACTGG -3′ and 5′-TCCACCACCCTGTTGCTGTA-3′.
Cfx96 real time quantitative pcr detection system
Manufactured by Bio-Rad
Sourced in China, United States
The CFX96 real-time quantitative PCR detection system is a laboratory instrument designed for precise DNA quantification and analysis. It utilizes real-time PCR technology to amplify and detect specific DNA sequences. The system provides accurate and reproducible results for a variety of applications, including gene expression analysis, pathogen detection, and DNA quantification.
Automatically generated - may contain errors
Lab products found in correlation
Trizol reagent, by Thermo Fisher Scientific (2 mentions)
Hiscript 3 1st strand cdna synthesis kit, by Vazyme (1 mentions)
Taq pro universal sybr qpcr master mix, by Vazyme (1 mentions)
Mlv transcriptase kit, by Thermo Fisher Scientific (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Sybr premix ex taq 2 2 kit, by Takara Bio (1 mentions)
Trizol reagent, by Takara Bio (1 mentions)
3 protocols using cfx96 real time quantitative pcr detection system
1
Quantifying Long Non-coding RNAs Expression
2
Real-time qPCR Analysis of CRC Cells
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Total RNA was extracted from HCT116 CRC cells using TRIzol® (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. cDNA was synthesized using the MLV transcriptase kit (Invitrogen; Thermo Fisher Scientific, Inc.). Fast SYBR Green master mix was used to determine the threshold cycle (Cq) value of each sample using a CFX96 real-time quantitative PCR detection system (Bio-Rad Laboratories, Inc.). GAPDH served as the gene used for normalization. The fold-changes were calculated using the relative quantification with 2−ΔΔCq (29 (link)). All reactions were performed in a 20 µl reaction volume in triplicate. The following PCR conditions were used: Initial denaturation at 95°C for 30 sec; followed by 40 cycles of 95°C for 5 sec and 60°C for 30–60 sec; and stage 3 was dissociation according to the manufacturer's protocol (cat. no. RR420; Takara Biotechnology Co., Ltd.).
The PCR primer sequences used were as follows: GAPDH forward, 5′-AAGGTCATCCCTGAGCTGAA-3′ and reverse, 5′-TGACAAAGTGGTCGTTGAGG-3′; G6PD forward, 5′-TGCATGAGCCAGATAGGCTG-3′ and reverse, 5′-GGTAGTGGTCGATGCGGTAG-3′; and PKM2 forward, 5′-ATGCAGCACCTGATAGCTCG-3′ and reverse, 5′-AGGCTCGCACAAGTTCTTCA-3′.
The PCR primer sequences used were as follows: GAPDH forward, 5′-AAGGTCATCCCTGAGCTGAA-3′ and reverse, 5′-TGACAAAGTGGTCGTTGAGG-3′; G6PD forward, 5′-TGCATGAGCCAGATAGGCTG-3′ and reverse, 5′-GGTAGTGGTCGATGCGGTAG-3′; and PKM2 forward, 5′-ATGCAGCACCTGATAGCTCG-3′ and reverse, 5′-AGGCTCGCACAAGTTCTTCA-3′.
3
Mouse Tissue RNA Extraction and RT-qPCR Analysis
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Total RNA was extracted from mouse tissues and cells with TRIzol reagent (TaKaRa, Dalian, China). Then, RT-qPCR was performed in a CFX96 real-time quantitative PCR detection system (Bio-Rad, Hercules, CA, USA) according to the instructions of SYBR Premix Ex Taq II (2×) kit (TaKaRa, Dalian, China). The relative expression levels of mRNAs and miRNAs were calculated using the 2−∆∆Ct method. ACTB was selected as the internal reference for mRNA, and U6 was selected as the internal reference for miRNA. The primer sequences used in this study are shown in Table S1 .
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