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J Vis Exp 2015 Mar 24;(97):e52650

High-throughput quantitative real-time RT-PCR assay for determining expression profiles of types I and III interferon subtypes.

Renn LA, Theisen TC, Navarro MB, Mane VP, Schramm LM, Kirschman KD, Fabozzi G, Hillyer P, Puig M, Verthelyi D, Rabin RL

Abstract

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-a subtypes, is limited by their shared sequence identity; likewise, the sequences of the type III IFN, especially IFN-¿2 and -¿3, are highly similar. This assay provides a reliable, reproducible, and relatively inexpensive means to analyze the expression of the seventeen interferon subtype transcripts.


Category: Journal Article
PubMed ID: #25867042 DOI: 10.3791/52650
PubMed Central ID: #PMC4401384
Includes FDA Authors from Scientific Area(s): Biologics Drugs
Entry Created: 2016-02-29
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