Primers and conditions used for RT-PCR
| Gene . | Primers . | Amplified product, bp . | Cycles . | Ta, °C . |
|---|---|---|---|---|
| ACTIN | S: 5′-atctggcaccacaccttctacaatgagctgcg-3′; AS: 5′-cgtcatactcctgcttgctgatccacatctgc-3′ | 838 | 40 | 63 |
| OCT4 | S: 5′-gggaaggtattcagccaaacg-3′; AS: 5′-ggttcgctttctctttcggg-3′ | 180 | 20 | 56 |
| BRACHYURY | S: 5′-cccgcgcactacacacccctcacc-3′; AS: 5′-ccttgggctgcggcgtcgtactg-3′ | 125 | 40 | 63 |
| SCL | S: 5′-atggtgcagctgagtcctcc-3′; AS: 5′-tctcattcttgctgagcttc-3′ | 331 | 40 | 63 |
| RUNX1B | S: 5′-cgtgcacatacattagtagcactacctttg-3′; AS: 5′-cttccacgaatcttgcttgcagaggttaag-3′ | 304 | 40 | 63 |
| RUNX1C | S: 5′-gaagtctgaacccagcatagtggtcagcag-3′; AS: 5′-gtggacgtctctagaaggattcattccaag-3′ | 231 | 40 | 63 |
| GATA-2 | S: 5′-agccggcacctgttgtgcaa-3′; AS: 5′-tgacttctcctgcatgcact-3′ | 244 | 30 | 58 |
| LMO2 | S: 5′-ggatcctgccggagagactatctc-3′; AS: 5′-gaattcagtgaacacctccgcaaa-3′ | 289 | 40 | 63 |
| C-MYB | S: 5′-ctcagacttggaaatgccttc-3′; AS: 5′-ccagtggtgtgagcagaaga-3′ | 406 | 40 | 63 |
| VE-CADHERIN | S: 5′-accggatgaccaagtacagc-3′; AS: 5′-acacactttgggctggtagg-3′ | 596 | 40 | 63 |
| FRIZZLED-3 | S: 5′-ccttggcctgaagatatgg-3′; AS: 5′-acaaggaggtgaacaactacg-3′ | 210 | 40 | 56 |
| FRIZZLED-4 | S: 5′-ccaacatggctgttgaaatg-3′; AS: 5′-tcacccaaccatttcctctc-3′ | 168 | 40 | 56 |
| FRIZZLED-5 | S: 5′-tgctaccagccgtccttcagt-3′; AS: 5′-ccatgccgaagaagtagaccag-3′ | 319 | 40 | 56 |
| WNT3A | S: 5′-ttcttactcctctgcagcctg-3′; AS: 5′-ctggatgccaatcttgatgc-3′ | 207 | 40 | 56 |
| WNT5 | S: 5′-cttcgcccaggttgtaattgaagc-3′; AS: 5′-ctgccaaaaacagaggtgttatcc-3′ | 273 | 40 | 56 |
| WNT10B | S: 5′-aatgcgaatccacaacaac-3′; AS: 5′-gggtctcgctcacagaagtc-3′ | 288 | 40 | 56 |
| GAPDH | S: 5′-gtcttctccaccatggagaaggct-3′; AS: 5′-catgccagtgagcttcccgttca-3′ | 277 | 40 | 50-63 |
| CYCLIN-D1 | S: 5′-gcggaggagaacaaacagat-3′; AS: 5′-tgtgaggcggtaggaca-3′ | 180 | 40 | 56 |
| Gene . | Primers . | Amplified product, bp . | Cycles . | Ta, °C . |
|---|---|---|---|---|
| ACTIN | S: 5′-atctggcaccacaccttctacaatgagctgcg-3′; AS: 5′-cgtcatactcctgcttgctgatccacatctgc-3′ | 838 | 40 | 63 |
| OCT4 | S: 5′-gggaaggtattcagccaaacg-3′; AS: 5′-ggttcgctttctctttcggg-3′ | 180 | 20 | 56 |
| BRACHYURY | S: 5′-cccgcgcactacacacccctcacc-3′; AS: 5′-ccttgggctgcggcgtcgtactg-3′ | 125 | 40 | 63 |
| SCL | S: 5′-atggtgcagctgagtcctcc-3′; AS: 5′-tctcattcttgctgagcttc-3′ | 331 | 40 | 63 |
| RUNX1B | S: 5′-cgtgcacatacattagtagcactacctttg-3′; AS: 5′-cttccacgaatcttgcttgcagaggttaag-3′ | 304 | 40 | 63 |
| RUNX1C | S: 5′-gaagtctgaacccagcatagtggtcagcag-3′; AS: 5′-gtggacgtctctagaaggattcattccaag-3′ | 231 | 40 | 63 |
| GATA-2 | S: 5′-agccggcacctgttgtgcaa-3′; AS: 5′-tgacttctcctgcatgcact-3′ | 244 | 30 | 58 |
| LMO2 | S: 5′-ggatcctgccggagagactatctc-3′; AS: 5′-gaattcagtgaacacctccgcaaa-3′ | 289 | 40 | 63 |
| C-MYB | S: 5′-ctcagacttggaaatgccttc-3′; AS: 5′-ccagtggtgtgagcagaaga-3′ | 406 | 40 | 63 |
| VE-CADHERIN | S: 5′-accggatgaccaagtacagc-3′; AS: 5′-acacactttgggctggtagg-3′ | 596 | 40 | 63 |
| FRIZZLED-3 | S: 5′-ccttggcctgaagatatgg-3′; AS: 5′-acaaggaggtgaacaactacg-3′ | 210 | 40 | 56 |
| FRIZZLED-4 | S: 5′-ccaacatggctgttgaaatg-3′; AS: 5′-tcacccaaccatttcctctc-3′ | 168 | 40 | 56 |
| FRIZZLED-5 | S: 5′-tgctaccagccgtccttcagt-3′; AS: 5′-ccatgccgaagaagtagaccag-3′ | 319 | 40 | 56 |
| WNT3A | S: 5′-ttcttactcctctgcagcctg-3′; AS: 5′-ctggatgccaatcttgatgc-3′ | 207 | 40 | 56 |
| WNT5 | S: 5′-cttcgcccaggttgtaattgaagc-3′; AS: 5′-ctgccaaaaacagaggtgttatcc-3′ | 273 | 40 | 56 |
| WNT10B | S: 5′-aatgcgaatccacaacaac-3′; AS: 5′-gggtctcgctcacagaagtc-3′ | 288 | 40 | 56 |
| GAPDH | S: 5′-gtcttctccaccatggagaaggct-3′; AS: 5′-catgccagtgagcttcccgttca-3′ | 277 | 40 | 50-63 |
| CYCLIN-D1 | S: 5′-gcggaggagaacaaacagat-3′; AS: 5′-tgtgaggcggtaggaca-3′ | 180 | 40 | 56 |
S indicates forward; and AS, reverse.