Results of multiplexing trials of qPCR markers. Average Ct values, with the range of copy numbers in parentheses, found across 24 replicates for markers run singly or combined in a multiplex reaction.
VCQA = Vector Control Quantitative Analysis; E = efficiency of amplification; CT = number of cycles.Quantitative analysis of DNA purity using VCQA employing qPCR.
aThe total volume of PCR reaction was 25 µl and the PCR conditions were as follows: 50°C for 2 min, 95°C for 10 min, followed by 40 cycles of 95°C, 15 s for denaturation and 60°C, 1 min for annealing
aUnderlined sequences are restriction enzymes. Bold sequence represents the histidine tag.* Negative PCR for B. ovis mutant strain.&Negative PCR for TT17573 strain.Primers for PCR used in this study.
