Quantitative RT-PCR analyses for miR-125b, miR-146a, miR-155, and RNU6 (used as normalization control) were performed using TaqMan miRNA assays with reagents, primers, and probes obtained from Qiagen. In brief, a stem loop primer was used for reverse transcription (30 min, 16°C; 30 min, 42°C; 5 min 85°C) followed by qPCR employing TaqMan probes and primers in a Bio-rad CFX96 Mastercycler. For assessing expression of SOCS-1, MyD88, b-actin, and the primary microRNAs (pri-miRs) mRNAs, cDNA was synthesized using a reverse transcription system (miScript II - Qiagem). qPCR was performed on the CFX96 Real-Time PCR Detection System (Bio-Rad Laboratories) as described (13 (link)). Primers and pri-miRNAs were purchased from Qiagen. Relative expression was calculated using the comparative threshold cycle (Ct) and expressed relative to control or WT (ΔΔCt method).
Cfx96 mastercycler
Manufactured by Bio-Rad
The CFX96 Mastercycler is a real-time PCR detection system designed for accurate and reliable nucleic acid amplification and detection. It features 96-well format, thermal cycling capabilities, and fluorescence detection to enable various real-time PCR applications.
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Lab products found in correlation
2 protocols using cfx96 mastercycler
1
Quantitative RT-PCR Analysis of miRNA
2
Quantification of miRNA and mRNA Levels
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Total RNA from cultured cells was isolated using the miRNeasy Mini Kit (Qiagen) according to the manufacturer’s instructions. Quantitative RT-PCR analyses for miR-21 and RNU6 (used as a normalization control) were performed using TaqMan miRNA assays with reagents, primers, and probes obtained from Qiagen. In brief, a stem loop primer was used for reverse transcription (30 min, 16°C; 30 min, 42°C; 5 min 85°C) followed by qPCR employing TaqMan probes and primers in a Bio-rad CFX96 Mastercycler. For assessing expression of Socs1, Myd88, Stat3, Il10, Chi3l3, Il6, Retnla, Nos2, Tnfa, Arg1, Mmp2, and actin, cDNA was synthesized using a reverse transcription system (miScript II—Qiagem). qPCR was performed on a CFX96 Real-Time PCR Detection System (Bio-Rad Laboratories) as described [9 (link)]. Primers were purchased from Integrated DNA technologies. Relative expression was calculated using the comparative threshold cycle (Ct) and expressed relative to control or WT (ΔΔCt method).
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