qPCR multiplex detection of microRNA and messenger RNA in a single reaction.

Khoury, S; Tran, N

qPCR multiplex detection of microRNA and messenger RNA in a single reaction.

Khoury, S

Tran, N

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Publisher:: PeerJ
Publication Type:: Journal Article
Citation:: PeerJ, 2020, 8, (6), pp. e9004
Issue Date:: 2020-01

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Field	Value	Language
dc.contributor.author	Khoury, S https://orcid.org/0000-0001-5631-7478
dc.contributor.author	Tran, N https://orcid.org/0000-0001-7747-2530
dc.date.accessioned	2020-09-02T03:38:54Z
dc.date.available	2020-03-26
dc.date.available	2020-09-02T03:38:54Z
dc.date.issued	2020-01
dc.identifier.citation	PeerJ, 2020, 8, (6), pp. e9004
dc.identifier.issn	2167-8359
dc.identifier.issn	2167-8359
dc.identifier.uri	http://hdl.handle.net/10453/142504
dc.description.abstract	Reverse Transcription-Quantitative PCR (RT-qPCR) is one of the standards for analytical measurement of different RNA species in biological models. However, current Reverse Transcription (RT) based priming strategies are unable to synthesize differing RNAs and ncRNAs especially miRNAs, within a single tube. We present a new methodology, referred to as RNAmp, that measures in parallel miRNA and mRNA expression. We demonstrate this in various cell lines, then evaluate clinical utility by quantifying several miRNAs and mRNA simultaneously in sera. PCR efficiency in RNAmp was estimated between 1.8 and 1.9 which is comparable to standard miRNA and random primer RT approaches. Furthermore, when using RNAmp to detect selected mRNA and miRNAs, the quantification cycle (Cq) was several cycles lower. This low volume single-tube duplex protocol reduces technical variation and reagent usage and is suitable for uniform analysis of single or multiple miRNAs and/or mRNAs within a single qPCR reaction.
dc.format	Electronic-eCollection
dc.language	eng
dc.publisher	PeerJ
dc.relation.ispartof	PeerJ
dc.relation.isbasedon	10.7717/peerj.9004
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	06 Biological Sciences, 11 Medical and Health Sciences
dc.title	qPCR multiplex detection of microRNA and messenger RNA in a single reaction.
dc.type	Journal Article
utslib.citation.volume	8
utslib.location.activity	United States
utslib.for	06 Biological Sciences
utslib.for	11 Medical and Health Sciences
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology
pubs.organisational-group	/University of Technology Sydney/Strength - CHT - Health Technologies
pubs.organisational-group	/University of Technology Sydney/Faculty of Engineering and Information Technology/School of Biomedical Engineering
utslib.copyright.status	open_access	*
dc.date.updated	2020-09-02T03:38:44Z
pubs.issue	6
pubs.publication-status	Published
pubs.volume	8
utslib.citation.issue	6

Abstract:

Reverse Transcription-Quantitative PCR (RT-qPCR) is one of the standards for analytical measurement of different RNA species in biological models. However, current Reverse Transcription (RT) based priming strategies are unable to synthesize differing RNAs and ncRNAs especially miRNAs, within a single tube. We present a new methodology, referred to as RNAmp, that measures in parallel miRNA and mRNA expression. We demonstrate this in various cell lines, then evaluate clinical utility by quantifying several miRNAs and mRNA simultaneously in sera. PCR efficiency in RNAmp was estimated between 1.8 and 1.9 which is comparable to standard miRNA and random primer RT approaches. Furthermore, when using RNAmp to detect selected mRNA and miRNAs, the quantification cycle (Cq) was several cycles lower. This low volume single-tube duplex protocol reduces technical variation and reagent usage and is suitable for uniform analysis of single or multiple miRNAs and/or mRNAs within a single qPCR reaction.

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http://hdl.handle.net/10453/142504