Table 1.
Primer information and PCR efficiency of the reference and target genes
| Gene name | Primer sequence (5′–3′) | Product size (bp) | Tm (°C) | Efficiency (%) | R 2 |
|---|---|---|---|---|---|
| CDS cloning | |||||
| PmFAR1 | AAGGAGACGATGGAGGTAAT | 1,638 | 56.8 | Not tested | Not tested |
| GCTATGGTTCCAATAGGGTA | 57.2 | ||||
| PmFAR2 | TTTGTCAACCACCGAGATCA | 1,640 | 56.7 | Not tested | Not tested |
| AAGCATCAGTTTTGAAAATACCTA | 55.8 | ||||
| Candidate reference gene | |||||
| RPL40 | F: TGGACAAGGCACCAAGCG | 226 | 57.6 | 97.65 | 0.9905 |
| R: ATGAAGGGTAGATTCTTTCTGGATA | 58.3 | ||||
| 18SrRNA | F: TCAAGACATGGTCGGAAGA | 242 | 57.9 | 103.54 | 0.9871 |
| R: GGGAGGCATTGCTGGTA | 57.2 | ||||
| β-actin | F: CATCCTGCGTTTGGATTTAG | 144 | 59.5 | 98.71 | 0.9926 |
| R: TCCAAAGCAACATAGCACAAT | 57.7 | ||||
| β-TUB | F: GTGGCGTTGTATGGTTCG | 123 | 56.8 | 99.34 | 0.9784 |
| R: CGGTATGGGTACTTTACTTATCTC | 57.6 | ||||
| ARF1 | F: AAGGAGGAGCCGCATCA | 203 | 60.3 | 102.18 | 0.9796 |
| R: GCATCCAAACCCACCATTA | 60.1 | ||||
| EF1-β | F: CCTGAGCCTATCGTTTGC | 240 | 58.8 | 97.56 | 0.9788 |
| R: CGCTTTCATGTCGGTTTC | 59.5 | ||||
| GAPDH | F: TCAAAACATCATCCCCGCAGCC | 129 | 60.3 | 100.24 | 0.9882 |
| R: CGTCAAGTCGACGACCGAGACGT | 59.2 | ||||
| Target genes for validation | |||||
| PmFAR1 | F: TATTCACGCAAATGGCAACG | 109 | 58.9 | 99.37 | 0.9849 |
| R: AGGCTTCTTCAGTAAATCACAGC | 58.5 | ||||
| PmFAR2 | F: TCATATTTCACGTCGCCGCTAG | 212 | 61.2 | 97.82 | 0.9943 |
| R: GGCTTCCCTCCAGTTCATCG | 60.7 |