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RT-qPCR cycle conditions for miRNAs and miRNAs amplification.
| Step | Temperature | Time |
|---|---|---|
| Reverse transcription | 55 °C | 10 min |
| Initial denaturation | 95 °C | 1 min |
| Denaturation | 95 °C | 10 s |
| Annealing /Extension | IR mRNA (54 °C) Irs-1 mRNA (56 °C), miRNA-155 (52 °C) miRNA 222 (57 °C) |
30 s |
| Melting curve | 60 °C | 30 s |
