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. Author manuscript; available in PMC: 2010 Apr 1.

Published in final edited form as: J Virol Methods. 2009 Jan 16;157(1):32–39. doi: 10.1016/j.jviromet.2008.12.005

Quantitative analysis of viral sequences. A. qRT-PCR was conducted on 10-fold serial dilutions of a plasmid harboring the complete PV1 genome using the PV1 primer set that amplifies a 171 bp product. The y-axis represents fluorescence and the x-axis represents cycle number. The PCR was carried out for 30 cycles. Graph shows amplification from a representative well. The amount of plasmid DNA and the corresponding average Ct from triplicate samples are indicated in the upper left. R² value obtained from the standard curve generated by plotting Ct values against plasmid copy number is shown. B. Total RNAs were extracted from uninfected or PV1-infected cells at an MOI of 10 at the indicated times post-infection and analyzed by qRT-PCR with primers for both PV1 and β-actin mRNAs. PV1 RNA copy number was determined from the standard curve obtained in A. The amount of PV1 RNA was normalized to the amount of β-actin mRNA using 1 hpi as reference. Values on the y-axis are expressed after log₁₀ transformation. The data represents viral RNA levels from a representative experiment. hpi: hours post infection.

Quantitative analysis of viral sequences. A. qRT-PCR was conducted on 10-fold serial dilutions of a plasmid harboring the complete PV1 genome using the PV1 primer set that amplifies a 171 bp product. The y-axis represents fluorescence and the x-axis represents cycle number. The PCR was carried out for 30 cycles. Graph shows amplification from a representative well. The amount of plasmid DNA and the corresponding average Ct from triplicate samples are indicated in the upper left. R² value obtained from the standard curve generated by plotting Ct values against plasmid copy number is shown. B. Total RNAs were extracted from uninfected or PV1-infected cells at an MOI of 10 at the indicated times post-infection and analyzed by qRT-PCR with primers for both PV1 and β-actin mRNAs. PV1 RNA copy number was determined from the standard curve obtained in A. The amount of PV1 RNA was normalized to the amount of β-actin mRNA using 1 hpi as reference. Values on the y-axis are expressed after log₁₀ transformation. The data represents viral RNA levels from a representative experiment. hpi: hours post infection.