Table 1 Primer list and amplification/sequencing strategies
PRIMER LIST | ||||
|---|---|---|---|---|
Gene | Primer | Forward primer (5'-3') | Reverse primer (5'-3') | (bp) |
AgAcp34A-1 | 69 ex | ATTGAACGAGCACCACCGC | ACAATGCAGAACCTTCGAC | 735 |
AgAcp34A-1 | 69 n | ACGCCAGGCTTGTACTCTC | CACTTATAAACTAGCTACC | 632 |
AgAcp34A-2/3 | 70 ex | ATTCCCGTAACTATCTTGC | TATAACTCACGGGCGATTC | 793 |
AgAcp34A-2/3 | 70 n | AATGTGTCCTTTCTGAACC | CTGCCCAATTAACCAATAG | 685 |
AgAcp34A-1/2/3 | exo1 (+) | TCGCCCTAGTGGCTGTTG | - | - |
AgAcp34A-2/3 | 3UTR (-) | - | TCGTCCATTCCCATCGCAG | - |
AgAcp34A-1/2 | exo2a (+) | TAGATAACAGACAGTTACC | - | - |
AgAcp34A-1/2 | exo2b (+) | ATGCCCATAAATACTTTAG | - | - |
AgAcp34A-1/2 | exo2a (-) | - | GTCAACAAGCCCTACAAGA | - |
AgAcp34A-3 | exo2c (+) | ATACCCAAMCTGCCTATGC | - | - |
AgAcp34A-3 | exo2d (+) | CTATGCGCCGGCAGGTTTC | - | - |
AgAcp34A-3 | exo2b (-) | - | ATTTTAGAAACCTGCCGGC | - |
AgAcp34A-1 | 69 C (-) | - | TCTATAGAYAGTATCTACG | - |
AgAcp34A-2/3 | 70 C (-) | - | AAATTGTTCATTGAGAGTC | - |
PCR STRATEGIES | ||||
Nested PCR strategy to selectively amplify AgAcp34A-2 and AgAcp34A-3 a | ||||
Gene | Portion | Forward primer (5'-3') | Reverse primer (5'-3') | (bp) |
AgAcp34A-2 | N terminal | 70 n (+)* | exo2a (-) | 326 |
| Â | C terminal | exo2a (+) | 3UTR (-)* | 276 |
AgAcp34A-3 | N terminal | 70 n (+)* | exo2b (-) | 296 |
| Â | C terminal | exo2c (+) | 3UTR (-)* | 289 |
Nested RT-PCR strategy to selectively amplify transcripts (cDNA) | ||||
Gene | Round | Forward primer (5'-3') | Reverse primer (5'-3') | (bp) |
AgAcp34A-1 | 1° round | exo1 (+) | 69 C (-) | 260 |
|  | 2° round | exo2a (+) | 69 C (-) | 170 |
AgAcp34A-2 | 1° round | exo1 (+) | 70 C (-) | 266 |
|  | 2° round | exo2a (+) | 70 c (-) | 176 |
AgAcp34A-3 | 1° round | exo1 (+) | 70 C (-) | 263 |
|  | 2° round | exo2c (+) | 70 C (-) | 189 |