Table 3 Quantitative RT-PCR primer sequences and reaction conditions for the target genes IGF2, IGF2R, and VEGF. Melting curve analyses were performed after each PCR and one sample from each triplicate was assessed by gel electrophoresis to confirm there was no contamination.
From: Expression and protein localisation of IGF2in the marsupial placenta
| Â | IGF2 | IGF2R | VEGF |
|---|---|---|---|
Fw primer 5' to 3' | CCTTTGTGGTGGGGAACTGGT | CACAGGAGGTGGAAATGGTGAA | GATGTCTATCAACGCAGCTACT |
Rv primer 5' to 3' | GGATGGGGTCTTCGCTGGGCA | CCCAGAGGCACTGAATAACTT | TGATGTTGTGCACCTCATAGGG |
Protocol | Â | Â | Â |
|  | 50°C 10 min | 50°C 10 min | 50°C 10 min |
|  | 95°C 15 min | 95°C 15 min | 95°C 15 min |
39 × | 95°C 30 sec | 95°C 30 sec | 95°C 30 sec |
|  | 60°C 20 sec | 55°C 20 sec | 55°C 20 sec |
|  | 72°C 1 min | 72°C 40 sec | 72°C 40 sec |
|  | 86°C 1 sec | 75°C 1 sec | 76°C 1 sec |
| Â | Plate read | Plate read | Plate read |