A total of 2 μg RNA from colon of syngeneic and allogeneic transplanted mice at day + 15 after alloSCT and complementary DNA, obtained using the RNeasy Mini Kit (Qiagen, Venlo, Netherlands) and the QuantiTect Reverse Transcription Kit (Qiagen) following the manufacturer’s instructions, were amplified (50°C, 2 min; 95°C, 10 min; 49 cycles of 95°C, 10 s; 60°C, 1 min) on DNA Engine Opticon (Bio-Rad, Hercules, CA, United States) using the TaqMan Gene Expression Master Mix (Life Technologies) and GPRC6a primers and probe from BioTez GmbH (Berlin, Germany). Data were analyzed with Opticon Monitor 3.1 analysis software (Bio-Rad) and the comparative CT Method (ΔΔCT Method).
Opticon monitor 3.1 analysis software
Manufactured by Bio-Rad
The Opticon Monitor 3.1 analysis software is a laboratory equipment product that provides real-time data analysis and monitoring capabilities for various experiments and applications. The software's core function is to collect, process, and display real-time data from compatible instruments and assays.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using opticon monitor 3.1 analysis software
1
Measuring GPRC6a Expression in Allogeneic Transplants
2
Quantitative Real-Time PCR Analysis
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RNA and cDNA were obtained using the RNeasy Mini Kit (QIAGEN, Venlo, Netherlands) and the QuantiTect Reverse Transcription Kit (QIAGEN) after manufacturer’s instructions. Real-time PCR amplification reaction (50°C for 2 min, 95°C for 10 min, 49 cycles of 95°C for 10 s, 60°C for 1 min) was performed on DNA Engine Opticon (BioRad, Hercules, CA, USA) using the TaqMan Gene Expression Master Mix (Life Technologies) and primers and probes (BioTez GmbH, Berlin, Germany) designed with the Primer Express 1.5 software (Life Technologies). Data were analyzed with the Opticon Monitor 3.1 analysis software (BioRad) and the comparative CT method (ΔΔCT method).
3
Protein Thermal Stability Assay
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Assay mixtures of 50 μl containing 0.6 mg/ml protein, 20 mM Bis-Tris propane, pH 7.0 and SYPRO Orange protein stain dye (Sigma). Melting curves were recorded with a temperature range of 20 to 95 °C, using a DNA Engine Opticon 3 real time PCR system (MJ Research) with Opticon Monitor 3.1 analysis software (BioRad).
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