Quantitation of viral RNAs by real-time PCR. Viral genomic DNA and RNAs were isolated from the same pool of purified mature capsids. RNAs with (+) or without (-) RNase treatment were reversely transcribed with oligo-dT and hexamers, and then used as templates in the real-time PCR with TaqMan probes. The calculated copy number of the mRNAs of E1Bsmall and fiber is shown as percentage relative to the copy number of the viral genomic DNA. The averaged cycle threshold (Ct) values of 5 replicates were used to determine the relative RNA copy number.