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

Described in this report is a qRT-PCR assay for the analysis of seventeen human IFN subtypes in a 384-well plate format that incorporates highly specific locked nucleic acid (LNA) and molecular beacon (MB) probes, transcript standards, automated multichannel pipetting, and plate drying. Determining...

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Detalles Bibliográficos
Autores principales: Renn, Lynnsey A., Theisen, Terence C., Navarro, Maria B., Mane, Viraj P., Schramm, Lynnsie M., Kirschman, Kevin D., Fabozzi, Giulia, Hillyer, Philippa, Puig, Montserrat, Verthelyi, Daniela, Rabin, Ronald L.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2015
Materias:
Immunology
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4401384/
https://www.ncbi.nlm.nih.gov/pubmed/25867042
http://dx.doi.org/10.3791/52650
Descripción
Sumario:Described in this report is a qRT-PCR assay for the analysis of seventeen human IFN subtypes in a 384-well plate format that incorporates highly specific locked nucleic acid (LNA) and molecular beacon (MB) probes, transcript standards, automated multichannel pipetting, and plate drying. Determining expression among the type I interferons (IFN), especially the twelve IFN-α subtypes, is limited by their shared sequence identity; likewise, the sequences of the type III IFN, especially IFN-λ2 and -λ3, are highly similar. This assay provides a reliable, reproducible, and relatively inexpensive means to analyze the expression of the seventeen interferon subtype transcripts.

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