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Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays
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 me...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Frontiers Media S.A.
2014
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4244598/ https://www.ncbi.nlm.nih.gov/pubmed/25505484 http://dx.doi.org/10.3389/fgene.2014.00413 |
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author | Le Carré, Joane Lamon, Séverine Léger, Bertrand |
author_facet | Le Carré, Joane Lamon, Séverine Léger, Bertrand |
author_sort | Le Carré, Joane |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4244598 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-42445982014-12-10 Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays Le Carré, Joane Lamon, Séverine Léger, Bertrand Front Genet Genetics 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. Frontiers Media S.A. 2014-11-26 /pmc/articles/PMC4244598/ /pubmed/25505484 http://dx.doi.org/10.3389/fgene.2014.00413 Text en Copyright © 2014 Le Carré, Lamon and Léger. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Genetics Le Carré, Joane Lamon, Séverine Léger, Bertrand Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays |
title | Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays |
title_full | Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays |
title_fullStr | Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays |
title_full_unstemmed | Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays |
title_short | Validation of a multiplex reverse transcription and pre-amplification method using TaqMan(®) MicroRNA assays |
title_sort | validation of a multiplex reverse transcription and pre-amplification method using taqman(®) microrna assays |
topic | Genetics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4244598/ https://www.ncbi.nlm.nih.gov/pubmed/25505484 http://dx.doi.org/10.3389/fgene.2014.00413 |
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