Table 3.
Primer names, sequence, amplification temperatures, and amplification efficiencies
| Genotyping | |||||
|---|---|---|---|---|---|
| Gene | Primer name | Forward primer | Reverse primer | A.T. | Enzyme |
| Vrn-H1 | UCW132_F/R | TGTTTTGCAAACTATTTGACCAG | TAGCGCTCATACCGTTCAAG | 59° | – |
| vrn-H1 | HvV1PromF2/R2 | ACTTCACCCAACCACCTGAC | CTGGCGGTTGATCTTGTTCT | 55° | – |
| Vrn-H2 | VRN-H2a_F/R | CATGAAACAGCAGCTCCAGA | TTTGCCTCTCTCTCCTGCAT | 59° | – |
| SNF2 | SNF_F/R | TTGGTACTTGAATGCCTGAAAA | ATGGCACAACTTGGATTTGA | 60° | – |
| Ppd-H1 | PPD/H1/F/R | CTGAGCCTGAAGAGGTCGAG | GTGGCGGGAGGTTATCTCT | 57° | BstUI |
| HvFT1 Prom. | HvFT1F/R1 | ATGGACATGGAACCTGCCACT | TGGTGATGATGAGTGTTGCCC | 55° | – |
| HvFT1 Intron | UCW133_F/R | TGCACACACTTAGCGCAGTA | GCAGACCGTGGAACTCAACT | 55° | RsaI |
| EBmac603 | EBmac603F/R | ACCGAAACTAAATGAACTACTTCG | TGCAAACTGTGCTATTAAGGG | 56° | – |
| Bmag914 | Bmag914F/R | GGGCAATATACAGTTCAACTC | ATGAACTGGAGGCAGTAAATA | 57° | – |
| qRT-PCRa | ||||
|---|---|---|---|---|
| Gene | Primer name | Forward primer | Reverse primer | Efficiency % |
| UCW118 | CNV_UCW118_F1/R1 | CAGTAAGGCGAACCATGTCATC | TGCGCACCAACACAGAACA | 99.9 |
| FT1_Prom. | CNV_FT1p_F2/R2 | CGGCCGAGTCTGTGTGATCT | GGCATAAATCCCGCCTCTTT | 99.9 |
| FT1_ATG | CNV_FT1p_F5/R4 | TGTTCTAAGAAGGAAGGAGAAATGG | GAAGGTCACCCTGAGGTTGGT | 99.8 |
| FT1_Ex1 | CNV_FT1_F2/R2 | CGTACGTACACAATCACCACTATCTAATG | GAGAGCCCGATCGTGCAT | 99.8 |
| FT1_Ex3 | CNV_FT1_F4/R3 | GCAGGTTGGTGACAGATATCCC | GGAAGAGCACGAGCACGAA | 89.3 |
| UCW123 | CNV_UCW123_F1/R1 | ACTGCAAGAGCTACAGCCTTCA | GTCACCGGCAGCAAGATCTAG | 99.9 |
| UCW120 | CNV_UCW120_F1/R1 | GCGACGACCAGTAAAAAATGC | CCGTTTCCGTGGATGGAA | 99.9 |
| SNF2 | CNV_SNF2_F1/R1 | ATTACCGCTCTGCTGTCGCGATTA | AAATGTGGCTCTGAAGGTGTTGGC | 93.4 |
aPCR program:(95° 20 s) × one cycle, (95° 3 s, 60° 30 s) × 40 cycles