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RNA decay measurement using Actinomycin D
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To determine the decay of viral mRNA species, HeLa cells infected with either Ad5 WT or ΔE1B were treated with 1 μg/ml Actinomycin D (Cayman Chemical, Cat#: 11421) at 24 hpi. RNA harvested using RLT buffer (from Qiagen RNA isolation kit) at 0, 1, 2, 4, 6, and 8 hours after treatment. RNA levels were quantified using RT-qPCR and normalized to 0 hours of Actinomycin D to determine RNA decay.
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