Total RNA was isolated using TRIzol kit. RT-qPCR analysis was conducted on a CFX96 Detection System with SYBR Green PCR Master Mix (Takara, Tokyo, Japan). The reaction conditions were 96°C for 1 min, 40 cycles of 96°C for 10 sec, 60°C for 5 sec, and 72°C for 30 sec. The primers were LINC01535, 5′-GGGCGGCAGGTCACTGACAC-3′ (forward) and 5′-GCCAGCAGCCGCTGGCTTAG-3′ (reverse); miR-146b-5p, 5′-TGACCCATCCTGGGCCTCAA-3′ (forward) and 5′-CCAGTGGGCAAGATGTGGGCC-3′ (reverse); TRIM2, 5′-TGGAGAAGGAAATGGGCATG-3′ (forward) and 5′-CTGCAACCACAACATGCACCA-3′ (reverse); GAPDH, 5′-CCAGCCGAGCCACATCGCTC-3′ (forward) and 5′-ATGAGCCCCAGCCTTCTCCAT-3′ (reverse); and U6, 5′-CTCGCTTCGGCAGCACA-3′ (forward) and 5′-AACGCTTCACGAATTTGCGT-3′ (reverse).
Cfx96 detection system
Manufactured by Takara Bio
Sourced in Japan, China
The CFX96 Detection System is a real-time PCR instrument designed for accurate and efficient gene expression analysis and quantification. It features a 96-well format and delivers reliable results with advanced thermal cycling and detection capabilities.
Automatically generated - may contain errors
Lab products found in correlation
Sybr green pcr master mix, by Takara Bio (1 mentions)
Minibest universal rna extraction kit, by Takara Bio (1 mentions)
Pcr master mix, by Takara Bio (1 mentions)
Trizol, by Thermo Fisher Scientific (1 mentions)
Reverse transcription reagent, by Takara Bio (1 mentions)
Trizol reagent, by Takara Bio (1 mentions)
Sybr premix ex taq, by Takara Bio (1 mentions)
3 protocols using cfx96 detection system
1
Quantitative Expression Analysis of ncRNAs
2
Quantitative RT-PCR Analysis of Gene Expression
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RNA was isolated from cultured cells using TRIzol (Invitrogen) in accordance with the manufacturer's instructions. Total RNA was isolated from fresh lung tumor and normal tissues using TaKaRa MiniBEST universal RNA extraction kit. The RNA quality was confirmed by Nanodrop spectrophotometry. Reverse transcription was performed with 1μg of total RNA following the manufacturer's instructions using an RT Kit (TakaRa, Dalian, China). Real-time PCR analysis was performed on an BioRad CFX96 Detection System using the PCR Master Mix (TakaRa, Dalian, China). The relative expression of each gene was normalized to beta-actin. The primers used for the quantitative RT-PCR are shown in supplementary Table 1 and supplementary Fig. 1 A.
3
Quantifying Gene Expression Using RT-qPCR
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Total RNA was isolated from cultivated cells with TRIzol reagent (TaKaRa, Dalian, China) and reverse-transcribed into complementary DNA (cDNA) using reverse transcription reagents (TaKaRa) as per the manufacturer’s instructions. On a Bio-Rad CFX96 detection system, SYBR Premix ExTaq (TaKaRa) was used to perform RT-qPCR. The 2−ΔΔCt method was used to calculate the fold changes in the gene expression. Primer were list in Supplementary Table 1
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