cDNA was generated from total RNA (0.8ug) using Quantitect Reverse Transcription kit (Qiagen, Cat#205310). RT-PCR analyses were performed using LightCycler 480 (Roche) in 10ul final volume, using 1ul of cDNA template. qPCR primers: 22 (link) and also new primers were designed (Universal Probe Library assay design tool, Roche, Supp.Table S10 ). Analyses were performed as previously described 22 (link).
Protocol detail
RT-PCR for Gene Expression Analysis
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Corresponding Organization :
Other organizations : Max Perutz Labs, University of Vienna, University of Manchester, Vienna Biocenter, University of St. Gallen, Stazione Zoologica Anton Dohrn
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Variable analysis
independent variables
- Reverse transcription using Quantitect Reverse Transcription kit (Qiagen, Cat#205310)
- MRNA expression levels measured by RT-PCR analyses using LightCycler 480 (Roche)
- Total RNA input (0.8 ug)
- CDNA template volume (1 ul)
- Final reaction volume (10 ul)
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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