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High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes
JoVE Journal
Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes

High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes

13,114 Views

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10:00 min

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March 24, 2015

DOI:

10.3791/52650-v

DOI:
10.3791/52650-v

•

10:00 min
March 24, 2015

•

3 Views
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1Center for Biologics Evaluation and Research,US Food and Drug Administration, 2Center for Drug Evaluation and Research,US Food and Drug Administration

Summary

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This protocol describes a high-throughput qRT-PCR assay for the analysis of type I and III IFN expression signatures. The assay discriminates single base pair differences between the highly similar transcripts of these genes. Through batch assembly and robotic pipetting, the assays are consistent and reproducible.

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QRT-PCRInterferon SubtypesType I InterferonsType III InterferonsLNA ProbesMolecular Beacon ProbesTranscript StandardsAutomated PipettingPlate DryingGene Expression Profiling

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Renn, L. A., Theisen, T. C., Navarro, M. B., Mane, V. P., Schramm, L. M., Kirschman, K. D., Fabozzi, G., Hillyer, P., Puig, M., Verthelyi, D., Rabin, R. L. High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes. J. Vis. Exp. (97), e52650, doi:10.3791/52650 (2015).

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