TABLE 2.
Standard curve data from a real-time quantitative reverse transcriptase PCR analysis of total RNA extracted from stimulated RAW cells, a macrophage like cell linea
| Target | ΔRg significance thresholdb | Log dilutions | Range of Ct valuesc | R2d | Slope |
|---|---|---|---|---|---|
| 18S rRNA | 0.2 | 10−3-10−7 | 7-26 | 0.997 | 3.7997 |
| TLR1 | 0.02 | 10−1-10−5 | 20-36 | 0.9805 | 3.444 |
| TLR2 | 0.04 | 10−1-10−5 | 18-36 | 0.9776 | 3.414 |
| TLR4 | 0.04 | 10−1-10−4 | 24-37 | 0.9891 | 3.719 |
| TLR5 | 0.02 | 10−1-10−3 | 29-38 | 0.9913 | 3.915 |
| TLR6 | 0.02 | 10−1-10−4 | 23-34 | 0.9561 | 3.219 |
| TLR9 | 0.03 | 10−1-10−4 | 24-34 | 0.9900 | 3.133 |
| MD2 | 0.04 | 10−1-10−4 | 19-38 | 0.9927 | 3.583 |
| MyD88 | 10−1-10−5 | 22-35 | 0.9829 | 3.106 | |
| TIRAP | 0.03 | 10−1-10−3 | 23-31 | 0.9320 | 3.435 |
| TNF-α | 0.03 | 10−2-10−6 | 17-38 | 0.9861 | 4.3687 |
| iNOS | 0.03 | 10−1-10−5 | 18-35 | 0.9805 | 3.495 |
a
To generate standard curves for the specific reactions for the various probes, total RNA extracted from stimulated RAW macrophages was serially diluted.
b
ΔRn is the change in the reporter dye level.
c
Ct is the threshold cycle value, the cycle at which the change in the reporter dye level detected passes the ΔRn.
d
R2 is the coefficient of regression. Regression analyses of the mean values of three or four replicate reverse transcriptase PCRs for the log10-diluted RNA were used.