Primers used to amplify rainbow trout target genes by real-time RT-PCR
| Gene . | Primer sequences . | Hybridization temperature (°C) . | Amplicon size (bp) . |
|---|---|---|---|
| EF1α | F: 5′-TCCTCTTGGTCGTTTCGCTG-3′ | 59 | 159 |
| R: 5′-ACCCGAGGGACATCCTGTG-3′ | |||
| GK | F: 5′-TGAAGGATCAGAGGTGGGTGATT-3′ | 59 | 253 |
| R: 5′-GAAGGTGAAACCCAGAGGAAGC-3′ | |||
| PK | F: 5′-CCATCGTCGCGGTAACAAGA-3′ | 59 | 158 |
| R: 5′-ACATAGGAAAGGCCAGGGGC-3′ | |||
| mPEPCK | F: 5′-GTTGGTGCTAAAGGGCACAC-3′ | 59 | 149 |
| R: 5′-CCCGTCTTCTGATAAGTCCAA-3′ | |||
| G6Pase 1 | F: 5′-CTCAGTGGCGACAGAAAGG-3′ | 55 | 77 |
| R: 5′-TACACAGCAGCATCCAGAGC-3′ | |||
| G6Pase 2 | F: 5′-TAGCCATCATGCTGACCAAG-3′ | 55 | 82 |
| R: 5′-CAGAAGAACGCCCACAGAGT-3′ | |||
| FBPase | F: 5′-GCTGGACCCTTCCATCGG-3′ | 59 | 182 |
| R: 5′-CGACATAACGCCCACCATAGG-3′ | |||
| FAS | F: 5′-TGATCTGAAGGCCCGTGTCA-3′ | 60 | 161 |
| R: 5′-GGGTGACGTTGCCGTGGTAT-3′ | |||
| CPT1A | F: 5′-TCGATTTTCAAGGGTCTTCG-3′ | 55 | 166 |
| R: 5′-CACAACGATCAGCAAACTGG-3′ | |||
| CPT1B | F: 5′-CCCTAAGCAAAAAGGGTCTTCA-3′ | 55 | 149 |
| R: 5′-CATGATGTCACTCCCGACAG-3′ |
| Gene . | Primer sequences . | Hybridization temperature (°C) . | Amplicon size (bp) . |
|---|---|---|---|
| EF1α | F: 5′-TCCTCTTGGTCGTTTCGCTG-3′ | 59 | 159 |
| R: 5′-ACCCGAGGGACATCCTGTG-3′ | |||
| GK | F: 5′-TGAAGGATCAGAGGTGGGTGATT-3′ | 59 | 253 |
| R: 5′-GAAGGTGAAACCCAGAGGAAGC-3′ | |||
| PK | F: 5′-CCATCGTCGCGGTAACAAGA-3′ | 59 | 158 |
| R: 5′-ACATAGGAAAGGCCAGGGGC-3′ | |||
| mPEPCK | F: 5′-GTTGGTGCTAAAGGGCACAC-3′ | 59 | 149 |
| R: 5′-CCCGTCTTCTGATAAGTCCAA-3′ | |||
| G6Pase 1 | F: 5′-CTCAGTGGCGACAGAAAGG-3′ | 55 | 77 |
| R: 5′-TACACAGCAGCATCCAGAGC-3′ | |||
| G6Pase 2 | F: 5′-TAGCCATCATGCTGACCAAG-3′ | 55 | 82 |
| R: 5′-CAGAAGAACGCCCACAGAGT-3′ | |||
| FBPase | F: 5′-GCTGGACCCTTCCATCGG-3′ | 59 | 182 |
| R: 5′-CGACATAACGCCCACCATAGG-3′ | |||
| FAS | F: 5′-TGATCTGAAGGCCCGTGTCA-3′ | 60 | 161 |
| R: 5′-GGGTGACGTTGCCGTGGTAT-3′ | |||
| CPT1A | F: 5′-TCGATTTTCAAGGGTCTTCG-3′ | 55 | 166 |
| R: 5′-CACAACGATCAGCAAACTGG-3′ | |||
| CPT1B | F: 5′-CCCTAAGCAAAAAGGGTCTTCA-3′ | 55 | 149 |
| R: 5′-CATGATGTCACTCCCGACAG-3′ |
R, reverse primer; F, forward primer
GenBank accession no. or sigenae accession no.: EF1α(elongation factor 1α), AF498320; GK (glucokinase), AF135403; PK (pyruvate kinase), AF 246146; mPEPCK (phosphoenolpyruvate carboxykinase, mitochondrial isoform), AF246149; G6Pase 1 (glucose 6-phosphatase 1), tcay0019b.d.18_3.1.s.om.8.1-1693; G6Pase 2 (glucose 6-phosphatase 2), AF120150; FBPase (fructose 1,6-bisphosphatase),AF333188; FAS (fatty acid synthase), tcab0001c.e.06_5.1.s.om.8; CPT1A (carnitine palmitoyltransferase 1 A), AF 327058; CPT1(carnitine palmitoyltransferase 1 B), AF606076