RT-PCR primers The annealing temperature of the RT-PCR is indicated; the rest of PCR conditions are indicated under Materials and Methods. The melting temperature and size of each amplicon was determined after each experiment.
Oligonucleotide | Annealing Temperature | Fragment Sizea | Fragment Temperature | |
|---|---|---|---|---|
| °C | bp | °C | ||
| E2F-1 (M96577)b | ||||
| Forward | 5′-AGGAGTTCATCAGCCTTTCCC-3′ | 60 | ||
| Reverse | 5′-CCCCAAAGTCACAGTCGAAGAG-3′ | 60 | 101 | 88.6 |
| Cyclin D1 (NM05305) | ||||
| Forward | 5′-TCCTCCTCTTCCTCCTCCTC-3′ | 55 | ||
| Reverse | 5′-TCAAGTGTGACCCAGACTGC-3′ | 55 | 124 | 90.7 |
| Cyclin D3 (BC011616) | ||||
| Forward | 5′-TTTTAGAGGGACCCCAATCC-3′ | 60 | ||
| Reverse | 5′-ACCAGCATTCCTTTTGAACG-3′ | 60 | 133 | 84.25 |
| Cyclin E (M73812) | ||||
| Forward | 5′-TCTGTAAAAGCAAACGCACG-3′ | 60 | 124 | |
| Reverse | 5′-AGCGGTAAGAAGCAGAGCAG-4′ | 60 | 88.3 | |
| Cdk2 (NM001798) | ||||
| Forward | 5′-ATGGACGGAGCTTGTTATCG-3′ | 60 | 165 | |
| Reverse | 5′-GGAGAGGGTGAGATTAGGGC-3′ | 60 | 87.2 | |
| Cdk4 (NM000075) | ||||
| Forward | 5′-CTGGTCGGCTTCAGAGTTTC-3′ | 60 | 120 | |
| Reverse | 5′-CCCGAAGTTCTTCTGCAGTC-3′ | 60 | 83.8 | |
| RNA 18S (X01117) | ||||
| Forward | 5′-GAGGTGAAATTCTTGGACCGG-3′ | 63 | ||
| Reverse | 5′-CGAACCTCCGACTTTCGTTCT-3′ | 63 | 94 | 82.7 |