Table 2. Primer sequences, target genes, and cycling conditions for SYBR green RT-PCR.
Target gene | Primer sequences (5’–3’) |
Reverse transcription | Primary denaturation |
Amplification (40 cycles) | Dissociation curve (1 cycle) |
Product size (bp) |
Accession no. | References | ||||
---|---|---|---|---|---|---|---|---|---|---|---|---|
Secondary denaturation | Annealing (optics on) |
Extension | Secondary denaturation | Annealing |
Final denaturation | |||||||
β-actin | F: CCACCGCAAATGCTTCTAAAC | 50°C 30 min |
94°C 5 min |
94°C 15 s |
51°C 30 s |
72°C 30 s |
94°C 1 min |
55°C 1 min |
94°C 1 min |
175 | NM205518 | [47] |
R: AAGACTGCTGCTGACACCTTC | ||||||||||||
ACC | F: AATGGCAGCTTTGGAGGTGT | 60.9°C 30 s |
60.9°C 1 min |
119 | NM_205505 | [48] | ||||||
R: TCTGTTTGGGTGGGAGGTG | ||||||||||||
CPT1 | F: CAATGAGGTACTCCCTGAAA | 57.5°C 30 s |
57.5°C 1 min |
337 | AY675193 | |||||||
R: CATTATTGGTCCACGCCCTC | ||||||||||||
PEPT1 | F: CCCCTGAGGAGGATCACTGTT | 60°C 30 s |
60°C 1 min |
205 | NM_204365 | [49] | ||||||
R: CAAAAGAGCAGCAGCAACGA | ||||||||||||
GLUT2 | F: CACACTATGGGCGCATGCT | 60°C 30 s |
60°C 1 min |
116 | NM_207178.1 | |||||||
R: ATTGTCCCTGGAGGTGTTGGTG |
β-actin, Beta actin; ACC, acetyl-CoA carboxylase; CPT1, carnitine acyltransferase I; PEPT1, peptide transporter 1; GLUT2, glucose transporter 2.