Table 5. Multiplex TaqMan qPCR assays with Liposcelis genomic DNA.
| Template conc. per reaction | Ct(SD) value of corresponding channel with genomic DNA | Ct(SD) value of corresponding channel with genomic DNA (performed after 8 months) | |||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|
| Orange | Yellow | Red | Crimson | Green | Orange | Yellow | Red | Crimson | Green | ||
| R2 | 0.999 | 0.999 | 0.999 | 0.999 | 0.999 | 0.999 | 0.999 | 0.999 | 0.999 | 0.999 | |
| Y | -3.319 | -3.364 | -3.384 | -3.444 | -3.433 | -3.370 | -3.178 | -3.368 | -3.266 | -3.388 | |
| Ex | 1.00 | 0.98 | 0.97 | 0.95 | 0.96 | 0.98 | 1.06 | 0.98 | 1.02 | 0.97 | |
| Ct(SD) values | 1 ng | 9.01(0.05) | 10.55(0.06) | 11.12(0.09) | 9.52(0.05) | 13.61(0.03) | 9.38(0.02) | 9.56(0.27)* | 11.39(0.15) | 10.33(0.17) | 12.68(0.20) |
| 100 pg | 12.29(0.05) | 14.03(0.06) | 14.56(0.04) | 13.08(0.02) | 17.02(0.11) | 12.75(0.04) | 11.66(0.02) | 14.72(0.11) | 13.67(0.04) | 16.07(0.06) | |
| 10 pg | 15.64(0.04) | 17.28(0.06) | 17.86(0.06) | 16.41(0.08) | 20.48(0.07) | 16.12(0.03) | 14.84(0.02) | 18.13(0.08) | 16.86(0.05) | 19.45(0.09) | |
| With single species | 20.06(0.08) | 21.64(0.13) | 20.76(0.11) | 27.09(0.17) | 23.64(0.41) | 24.29(0.25) | 18.54(0.19) | 19.68(0.02) | 25.67(0.13) | 22.65(0.21) | |
| With all five species | 23.37(0.32) | 21.85(0.15) | 20.67(0.15) | 31.53(1.23) | 23.37(0.33) | 22.27(0.05) | 17.57(0.02) | 19.19(0.01) | 28.34(0.87) | 22.30(0.12) | |
| NTC | - | - | - | - | - | - | - | - | - | - | |
Average Ct values using genomic DNA of Liposcelis brunnea, L. bostrychophila, L. decolor, L. obscura and L. pearmani with primer and probe sets BruCo5F/5R/5P, BosCo8F/8R/8P, DecCo11F/11R/11P, ObsCo12F/12R/12P and PeaCo14F/14R/14P, respectively.
*Due to error in fluorescence, this value has not been included in analysis; R2, linear correlation; Y, slope; Ex, reaction efficiency; SD, standard deviation; the Ct values are average of three replicates; Orange, yellow, red, crimson and green color corresponding to L. obscura, L. brunnea, L. bostrychophila, L. pearmani and L. decolor, respectively.