Development of Strand-Specific Real-Time RT-PCR to Distinguish Viral RNAs during Newcastle Disease Virus Infection

Figure 2

The development of a real-time quantitative PCR assay for absolute quantification. (a) Melting curve, (b) linear regression equation. The amount of plasmid T-NP was converted into copy number via the following equation: copy number = (amount of T-NP × 6.022 × 10²³)/(length of T-NP × 10⁹ × 660). The linearity of the standard curve was retained when the plasmid concentration was in the range of 5.5 × 10²–1.1 × 10⁹ copies/μL. The obtained linear regression equation was ( = 0.998). The amplification efficiency was 101%.

(a)

(b)