RNA was first isolated using the RNeasy Mini Kit (QIAGEN) and then reverse transcribed using the High-Capacity RNA-to-cDNA kit (Thermo Fisher Scientific) according to the manufacturers’ instructions. RT-qPCR was performed using the SsoAdvanced Universal Supermix (Biorad) with Quantitect primers (TPP1 only) or previously validated primers20 (link),27 (link). Primer sequences available upon request.
Protocol detail
Quantifying Messenger RNA Expression
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Corresponding Organization :
Other organizations : The Francis Crick Institute, Children's Medical Research Institute, University of Sydney
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Variable analysis
independent variables
- RNA isolation using the RNeasy Mini Kit (QIAGEN)
- Reverse transcription using the High-Capacity RNA-to-cDNA kit (Thermo Fisher Scientific)
- RT-qPCR results
- Manufacturers' instructions for the RNeasy Mini Kit, High-Capacity RNA-to-cDNA kit, and SsoAdvanced Universal Supermix (Biorad)
- No positive or negative controls were explicitly mentioned in the provided information.
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