Table 2.
Primers used in this study for detection of Glu-1Bx alleles
| PS | Gene | Detection region | Marker name | Type of Primer | Sequence (5’-3’) | AT | Agarose gel (%) | Reference | |
|---|---|---|---|---|---|---|---|---|---|
| i-MAX II | Ex Taq | ||||||||
| PS1 | Glu-1Bx7/17 | CDS | MHBx717-F | SNP | CACTGAGATGGCTAAGCGCC | 66℃ | 66℃ | 1% | this study |
| MHBx717-R | GATCTTGTTGCCCTTGTCC | ||||||||
| PS2 | Glu-1Bx7 OE | Promoter | MAR-F | Indel (+ 43 bp) | CCTCAGCATGCAAACATGCAGC | 66℃ | 68℃ | 1% | [23] |
| MAR-R | CTGAAACCTTTGGCCAGTCATGTC | ||||||||
| PS3 | Glu-1Bx7 OE | Junction | TaBAC1215C06-F517 | Duplication | ACGTGTCCAAGCTTTGGTTC | 60℃ | 68℃ | 1% | [33] |
| TaBAC1215C06-R964 | GATTGGTGGGTGGATACAGG | ||||||||
| PS4 | Glu-1Bx7 OE | Junction | TaBAC1215C06-F24671 | Duplication | CCACTTCCAAGGTGGGACTA | 60℃ | 68℃ | 1% | [33] |
| TaBAC1215C06-R2551 | TGCCAACACAAAAGAAGCTG | ||||||||
| PS5 | Glu-1Bx20 | Promoter | MHBx-185-F | Indel (+ 185 bp) | GATAAGGCCAACGAGAGAAGAA | 60/66℃ | 59℃ | 1% | this study |
| MHBx-185-R | GATCTTAGTAAATCCGCGTCAAATAG | ||||||||
| PS6 | Glu-1Bx20 | CDS | cauBx752-F | Indel (+ 185 bp) | AGGGGCAGGGAAGAAACACT | 66℃ | 66℃ | 1% | [31] |
| cauBx752-R | CCAGGCAACACAAATCCATG | ||||||||
| PS7 | Glu-1Bx7* | CDS | Bx7F | Indel (-18 bp) | CAACTTCTTCACAGCAGT | 60℃ | 66℃ | 3% | [35] (modified) |
| Bx7R | TGCGCCTTTGCCACCTTTAG | ||||||||
| PS8 | Glu-1Bx17 | CDS | cauBx642-F | Indel (-108 bp) | GGGCAATCGGGGTACTTCC | 60℃ | 59℃ | 1% | [31] |
| cauBx642-R | CCCTTGTCTTGGCTGTTGTC | ||||||||
| PS9 | Glu-1Bx6 | CDS | BX7-F | InDel (+ 15 bp) | CACTGAGATGGCTAAGCGCC | 60℃ | 66℃ | 3% | [32] |
| BX7-R | GCCTTGGACGGCACCACAGG | ||||||||
| PS10 | Glu-1Bx6 | CDS | MHBx6-F | SNP | CACTGAGATGGCTAAGCGCC | 67℃ | 68℃ | 1% | this study |
| MHBx6-R | GATCTTGTTGCCCTTGTTC | ||||||||
| PS11 | Glu-1Bx13 | Promoter | MHpro13-F | Indel (-54 bp) | GGGCTTTAGGAGAGATGGTTTAT | 60℃ | 59℃ | 1% | this study |
| MHpro13-R | GCCTATGAAGAAAGCGTGAGA | ||||||||
| PS12 | Glu-1Bx13 | CDS | MHBx13-F | SNP | GGCACGAGGCAATACGAGCG | 66℃ | 66℃ | 1% | this study |
| MHBx13-R | TGATCTTGTTGCTCTTGTCT | ||||||||
| PS13 | Glu-1Bx20 | CDS | MHBx20-F | SNP | GTGTACTACCCAACTTCTCT | 66℃ | 68℃ | 1% | this study |
| MHBx20-R | CCTGGCTGTTGCTCATGTCT | ||||||||
| PS14 | Glu-1Bx14 (−) | CDS | MHBx14(-)-F | Indel (-18 bp) | TACTACCCAACCTCTCCACA | 64℃ | 66℃ | 3% | this study |
| MHBx14(-)-R | CTGTTGCTCTTGTGCTGATTG | ||||||||
PS Primer set, CDS Coding sequence, AT Annealing temperature, new Newly developed marker