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Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays.

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journal contribution
posted on 2014-11-26, 00:00 authored by J Le Carré, Severine LamonSeverine Lamon, B Léger
Since the discovery of microRNAs (miRNAs), different approaches have been developed to label, amplify and quantify miRNAs. The TaqMan(®) technology, provided by Applied Biosystems (ABIs), uses a stem-loop reverse transcription primer system to reverse transcribe the RNA and amplify the cDNA. This method is widely used to identify global differences between the expression of 100s of miRNAs across comparative samples. This technique also allows the quantification of the expression of targeted miRNAs to validate observations determined by whole-genome screening or to analyze few specific miRNAs on a large number of samples. Here, we describe the validation of a method published by ABIs on their web site allowing to reverse transcribe and pre-amplify multiple miRNAs and snoRNAs simultaneously. The validation of this protocol was performed on human muscle and plasma samples. Fast and cost efficient, this method achieves an easy and convenient way to screen a relatively large number of miRNAs in parallel.

History

Journal

Frontiers in genetics

Volume

5

Pagination

413-1

Location

Lausanne, Switzerland

eISSN

1664-8021

Language

eng

Publication classification

C Journal article, C1 Refereed article in a scholarly journal

Copyright notice

2014, Frontiers Research Foundation

Publisher

Frontiers Research Foundation