TABLE 1.

Average CT values for different real-time RT-PCR experiments using PUUV strain UME

No. of copiescDNAaAvg CT value for expt no.:Mean CV (%)c
12345678910Intra-assayInterassay
2 × 106P18.9319.0219.3219.2219.1919.3019.3318.9919.0019.340.210.85
GNDbNDNDNDNDNDNDNDNDNDNDND
2 × 105P22.2722.4923.0422.8422.5222.9422.5822.3322.2922.940.251.30
GNDNDNDNDNDNDNDNDNDNDNDND
2 × 104P25.6425.7626.3525.9925.7426.1025.9125.5025.5526.090.201.05
G24.9325.1025.1825.2625.2625.2625.2025.0824.8425.040.120.58
2 × 103P28.6529.0629.7129.2229.1229.3229.3128.7728.9129.500.281.12
G28.3628.4628.5828.7028.7128.7528.6028.5928.5228.400.260.46
2 × 102P32.0832.2233.1632.8232.4232.4932.6832.1732.1632.910.421.13
G31.8831.8431.8031.9532.2732.0432.1032.0332.0032.060.370.43
2 × 101P36.2235.5735.5635.5835.7035.3236.3735.6035.7735.931.450.91
G34.8935.5134.9735.4635.4834.8034.7235.0335.1135.621.020.94
2 × 100PNDNDNDNDNDNDNDNDNDNDNDND
G38.6637.4338.750.0037.9237.660.0037.2138.350.001.221.59
  • a P, plasmid cDNA; G, genomic cDNA.

  • b ND, not determined.

  • c Consistency (intra-assay CV) and reproducibility (interassay CV) for the real-time RT-PCR method. The copy numbers were spectrophotometrically calculated for the template RNA used in the plasmid reactions but only approximated for the genomic PUUV RNA/cDNA.