RNA was isolated from TG and converted into cDNA as described (9 (link)). Relative gene expression was calculated by the standard 2−ΔΔCt method, standardized reference genes, and normalized to WT UI controls following semi-quantitative real time PCR using a CFX Connect thermocycler (Bio-Rad, Hercules, CA). Viral transcripts were amplified using iTaq supermix (Biorad) with real-time primers from Integrated DNA Technologies (Coralville, IA). Primer sequences are listed in Supplemental Table I . PrimePCR technology (Bio-Rad) was utilized for ISG transcript expression studies according to the manufacturer’s instructions. Profiles of transcript expression represented by the cluster image map (or heat map) data supplement were generated using the National Cancer Institute’s CIMminer tool freely accessible online.
Protocol detail
Quantifying Viral Transcript Abundance
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Other organizations : University of Oklahoma Health Sciences Center
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Variable analysis
independent variables
- None explicitly mentioned
- Relative gene expression
- Viral transcripts
- WT UI (wild-type uninduced) controls
- Positive control: None mentioned
- Negative control: WT UI (wild-type uninduced) controls
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