Fibroblasts are a site of murine cytomegalovirus lytic replication and Stat1-dependent latent persistence in vivo - Nature Communications

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Fibroblasts are a site of murine cytomegalovirus lytic replication and Stat1-dependent latent persistence in vivo - Nature Communications
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Study sheds light on a herpesvirus lurking in the connectivetissue NatureComms

BAC copies contained 1 ng carrier RNA . Cycling conditions were as follows: enzyme activation, 2 min 95 °C, followed by 50 cycles of denaturation for 10 s at 95 °C, annealing for 20 s at 57 °C and extension for 30 s at 72 °C. Specificity of amplified sequences was validated by inspection of melting curves. RT-qPCR reactions prepared from equal amount of sample RNA but with the omission of reverse transcriptase served as controls for the absence of carry-over viral DNA.

To ensure detection of even minimal amounts of virusfor 30 min and then incubated at 37 °C for another 30 min. Afterwards, the inoculum was removed and the cells overlaid with medium containing 0.75% methylcellulose . After 4 days at 37 °C, the plates were examined for the presence of plaques under an inverted microscope.at an MOI of 0.1 for an hour at 37 °C.

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