Table 2.
Target gene | Primers sequences | Product size (bp) | Reverse transcription | Primary denaturation |
Amplification (40 cycles) | Dissociation curve (1 cycle) |
Reference | ||||
---|---|---|---|---|---|---|---|---|---|---|---|
Secondary denaturation | Annealing (Optics on) |
Extension | Secondary denaturation | Annealing | Final denaturation | ||||||
β-actin (EU887951) |
CCACACAGTGCCCATCTACGA CCACGCTCTGTCAGGATCTTCA | 89 |
50 °C 30 min |
94 °C 5 min |
94 °C 15 s |
62 °C 30 s |
72 °C 30 s |
94 °C 1 min |
62 °C 1 min |
94 °C 1 min |
Qiang et al. 2014 |
TGF-β |
GTTTGAACTTCGGCGGTACTG TCCTGCTCATAGTCCCAGAGA |
80 |
50 °C 30 min |
94 °C 5 min |
94 °C 15 s |
60 °C 30 s |
72 °C 30 s |
94 °C 1 min |
60 °C 1 min |
94 °C 1 min |
Standen et al. 2016 |
IL-1β |
TGGTGACTCTCCTGGTCTGA GCACAACTTTATCGGCTTCCA | 86 |
50 °C 30 min |
94 °C 5 min |
94 °C 15 s |
62 °C 30 s |
72 °C 30 s |
94 °C 1 min |
62 °C 1 min |
94 °C 1 min |
Standen et al. 2016 |
IL-10β |
CTGCTAGATCAGTCCGTCGAA GCAGAACCGTGTCCAGGTAA |
94 |
50 °C 30 min |
94 °C 5 min |
94 °C 15 s |
60 °C 30 s |
72 °C 30 s |
94 °C 1 min |
60 °C 1 min |
94 °C 1 min |
Standen et al. 2016 |
TNF-α (AY428948) |
CCAGAAGCACTAAAGGCGAAGA CCTTGGCTTTGCTGCTGATC |
82 |
50 °C 30 min |
94 °C 5 min |
94 °C 15 s |
60 °C 30 s |
72 °C 30 s |
94 °C 1 min |
60 °C 1 min |
94 °C 1 min |
Standen et al. 2016 |