Table 1 Primer and probe designs and real-time PCR conditions
Target | Forward primer | Reverse primer | UPL probe |
|---|---|---|---|
GAPDH | 5′-ggg-ttc-cta-taa-ata-cgg-act-gc-3′ | 5′-cca-ttt-tgt-cta-cgg-gac-ga-3′ | No. 52 |
FlaA | 5′-gtt-tct-ggg-gcc-aat-gta-ag-3′ | 5′-gcc-ttt-ggc-aaa-acc-aat-ag-3′ | No. 120 |
TNF-α | 5′-tct-tct-cat-tcc-tgc-ttg-tgg-3′ | 5′-ggt-ctg-ggc-cat-aga-act-ga-3′ | No. 49 |
IL-1β | 5′-ttg-acg-gac-ccc-aaa-aga-t-3′ | 5′-gaa-gct-gga-tgc-tct-cat-ctg-3′ | No. 26 |
IFN-γ | 5′-atc-tgg-agg-aac-tgg-caa-aa-3′ | 5′-ttc-aag-act-tca-aag-agt-ctg-agg-ta-3′ | No. 21 |
Fas | 5′-tgc-aga-cat-gct-gtg-gat-ct-3′ | 5′-ctt-aac-tgt-gag-cca-gca-agc-3′ | No. 34 |
FasL | 5′-acc-ggt-ggt-att-ttt-cat-gg-3′ | 5′-agg-ctt-tgg-ttg-gtg-aac-tc-3′ | No. 21 |
Cycle | Temperature (°C) | Duration (seconds) | Cycles |
|---|---|---|---|
Thermal cycler conditions | |||
Pre-incubation | 95 | 600 | 1 |
Amplification | 95 | 10 | 45 |
60 | 30 | ||
72 | 1 | ||
Cooling | 40 | 30 | 1 |
