Figure 6

Internal control target regions and design – both AIC-1 and AIC-2 were artificially synthesized. (a) Artificial internal control AIC-1 contains target for primer RT-F1, RT-R1, RT-R3 and a unique probe IC-P1; (b) artificial internal control AIC-2 contains target for primer IC-PF2, IC-PR2 and a unique probe IC-P1; (c) host internal control HIC is an internal transcribed spacer sequence of host genome (Lolium rigidum) contains target for primers IC-PF2, IC-PR2 and probe IC-PLT. AIC-1 can be detected by using the same primer as for R. toxicus—no need to add another set of primer but require an additional unique probe to be detected by different channel of qPCR. AIC-2 can be detected by using the same primer set as to detect host genome—no need to add any additional primer set to detect AIC-2 but require an additional unique probe to be detected by different qPCR channel. An additional primer and probe is required for HIC.