Table 1.
Primers Used for Cloning, RT-PCR, QPCR, and Bisulfite Sequencing Analyses of High-Affinity Potassium Transporter Genes in Bread Wheat
| S. No. | Sequence | Annealing temp. (°C) | Product size (bp) | Usage |
|---|---|---|---|---|
| HKT1;4 gene | ||||
| 1 | Forward primer: ATTCAGGCAACACCTAATCATGC | 56 | 473 | RT-PCR |
| Reverse primer: GCATCACAAGAATGAGGATGAGC | 581 | Cloning | ||
| 2 | Forward primer: TTTCTGTTCCAGGTACCTGCCTCCATACA | 49 | 384 | Bisulfite sequencing |
| Reverse primer: ARAARCCCCCATTTCCATCCRCACTRC | ||||
| 3 | Forward primer: ACCTCGCCATCTTCATCATC | 56 | 199 | qPCR |
| Reverse primer: GCTTCCATGAAGGAAACCAA | ||||
| Actin gene | ||||
| 4 | Forward primer: TGGGATGCCACCAAAGAC | 56 | 380 | RT-PCR and qPCR |
| Reverse primer: TGATACGCAAATGTTGAGC | ||||
| Ferredoxin-NADP(H) oxidoreductase (TaFNRII) gene | ||||
| 5 | Forward primer: CAGTGATCTTCACTTCTGAAC | 56 | 200 | RT-PCR and qPCR |
| Reverse primer: CGAGGACAAGAACGGGAAG | ||||
| HKT2;1 gene | ||||
| 6 | Forward primer: TATGTGATGAGTCGCAGCTTGAA | 56 | 316 | qPCR |
| Reverse primer: GCAACAAGAGGCCTGAATTCTTT | ||||
| 7 | Forward primer: TTYAATTYAGYYAAGAATGTAYAGAG | 49 | 254 | Bisulfite sequencing |
| Reverse primer: AARAACCATARTTTCATTTARARRCAC | ||||
| HKT2;3 gene | ||||
| 8 | Forward primer: TGAAGCCAAGCAACCCTAAC | 56 | 178 | qPCR |
| Reverse primer: CCAAGCAGGGAAACAAACAT | ||||
| 9 | Forward primer: GAATTATTTGGTGTTTTATTTTTYGGTTT | 51 | 369 | Bisulfite sequencing |
| Reverse primer: ACACRATAACCRATATAACTCTACTATC | ||||
HKT, high-affinity potassium transporter; RT-PCR, reverse transcription-polymerase chain reaction; qPCR, quantitative PCR.