Table 3.

The effect of adding different amounts of non-target (SED, 331 bp) amplicons to the RB1 assay. The effect is shown for different polymerase types (Ex Taq HS or Taq polymerase), polymerase amounts (0.5 U, 1 U or 2 U) and annealing/extension times (1x, 2x or 3x). Data are presented as ΔCq ± standard deviation (n = 3). ΔCq = Cq(added non-target amplicons) – Cq(control). ΔCq values above zero indicate partially impaired amplification. ND – no detected amplification.

qPCR parameters			Amount of non-target 331 bp amplicon
DNA polymerase	Polymerase amount	Annealing/extension time	0.5 μg	1 μg	1.5 μg	2 μg
Ex Taq HS	0.5 U	1x	−0.05 ± 0.03	–	ND	–
Ex Taq HS	1 U	1x	−0.44 ± 0.01	0.35 ± 1.06	8.79 ± 1.77	ND
Ex Taq HS	2 U	1x	−0.11 ± 0.12	–	0.40 ± 0.06	–
Ex Taq HS	1 U	2x	−0.33 ± 0	–	2.19 ± 0.84	–
Ex Taq HS	1 U	3x	−0.12 ± 0.04	–	0.18 ± 0.36	–
Taq	0.5 U	1x	4.90 ± 0.36	–	ND	–
Taq	1 U	1x	0.58 ± 0.40	6.80 ± 0.26	ND	ND
Taq	2 U	1x	−0.10 ± 0.02	–	0.22 ± 0.02	–
Taq	1 U	2x	0.25 ± 0.03	–	14.09 ± 2.20	–
Taq	1 U	3x	0.36 ± 0.03	–	0.97 ± 0.21	–