Table 2.
Primer sequences used for amplification of reference genes and GOI, amplicon length, melting temperature, and PCR efficiency.
| Gene abbreviation | Primer sequence (forward/reverse primer) | Amplicon length | Tm | Efficiency |
|---|---|---|---|---|
| ACT7 | 5′-AGACAACACGGCCTGGATAG-3′5′-TGAAGAGCACCCGGTTTTAC-3′ | 130 bp | 81°C | 104% |
| PROF | 5′-CCCAATTTCCCTCAGTTGAA-3′5′-TTGGTCCCACCACAGTGTAA-3′ | 104 bp | 81°C | 83.5% |
| PP2AA3 | 5′-TGCTTACCCTAGTGCCTCTGA-3′5′-TTCCCAAATTTGTAGCAGCA-3′ | 113 bp | 80°C | 96.2% |
| GAPDH | 5′-GGTGAGAAACCCGTCACTGT-3′5′-CTTTGCACCACCCTTCAAGT-3′ | 104 bp | 85.5°C | 82.5% |
| αTUB | 5′-GCACTTACCGCCAACTCTTC-3′5′-TTACTCGGTCAAGGCAAAGG-3′ | 126 bp | 82.5°C | 86.8% |
| βTUB | 5′-TTTCCCGGTCAACTCAACTC-3′5′-ACTGTGAGGGCTCGGTATTG-3′ | 134 bp | 85.5°C | 95.5% |
| Clath | 5′-TGCTTCCGCCATCTACTTTT-3′5′-TCCCAAGTTCCTTTGTTTGC-3′ | 128 bp | 80.5°C | 91% |
| CYP5 | 5′-CAGTGCCAAAAACAGCAGAA-3′5′-GGTGAAGTCACCTCCCTGAA-3′ | 143 bp | 82°C | 92.4% |
| EF1α | 5′-AAGCCAGGTATGGTCGTCAC-3′5′-GACATTGTCTCCGGGAAGAG-3′ | 108 bp | 85.5°C | 96.2% |
| PP2AA2 | 5′-CATGGGCTCAGAAATCACCT-3′5′-ATTGGTCAACGATGGGGATA-3′ | 128 bp | 82.5°C | 93.9% |
| SAND | 5’-TGCTTACACCACAAGGCAAG-3′5′-GAAGCAGCATGTCATCAGGA-3′ | 149 bp | 82°C | 104.9% |
| TIP41 | 5′-GTTGGGTGCGCATCTCTAAT-3′5′-AGCTCCGGCAGCTTTTACTT-3′ | 100 bp | 83.5°C | 93.2% |
| TUB ß9 | 5′-GCGATTCATTGCAAGGTTTT-3′5′-CATCATTCGATCGGGGTATT-3′ | 113 bp | 84°C | 98.4% |
| UBC | 5′-GGTGGAGTTTTTCAGCTTGC-3′5′-TCAAGGCAAATCTCTCCTGTC-3′ | 125 bp | 79.5°C | 99.1% |
| ACT2 | 5′-AGGATCTTCAGCCCCTTGTT-3′5′-ACCATTGTCTGGCAGCCTAC-3′ | 119 bp | 84°C | 102% |
| bHLH | 5′-GCAAAGTGGCTAAAGCTTCC-3′5′-CATGGGCTTCTTCCAAGTGT-3′ | 146 bp | 79.5°C | 107% |
| UBQ10 | 5′-GCGTTTGTTCATTGCTTCAA-3′5′-ATGGTGTCCGAGATTTCCAC-3′ | 177 bp | 81°C | 99.3% |
| UBC9 | 5′-CGGTCCTGTAGCGGAAGATA-3′5′-AGTCCTGAATGCAACCTTGG-3′ | 145 bp | 82.5°C | 90% |
| AOC | 5′-CGACAGGAGGTTAGGAGCTG-3′5′-CGCATCCAGTAGCTTCATCA-3′ | 115 bp | 86°C | 101% |
| MYC2 | 5′-GGCTAATGGCGGTAATGAAA-3′5′-GTCAAGGCTAATCGGAGCTG-3′ | 142 bp | 83.5°C | 90% |
| PAL | 5′-CATGGACAACACTCGTTTGG-3′5′-TTACGAGCTCGGAGAATTGG-3′ | 71 bp | 82°C | 96.6% |
| C4H | 5′-CAAGCTTCCACCTGGACCTA-3′5′-GCCCATACGGAGAAGCAATA-3′ | 130 bp | 85°C | 90.5% |
| 4CL | 5′-ATCAATGAGCCCGATGTCTC-3′5′-CTTAACGATCCGGAAGCAAC-3′ | 75 bp | 84°C | 90% |
The genes discarded from the stability analyses are underlined. Gene abbreviations are explained in Table 1.