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miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample

BACKGROUND: MicroRNAs (miRNAs) are small endogenous non-coding interfering RNA molecules regarded as major regulators in eukaryotic gene expression. Different methods are employed for miRNA expression profiling. For a better understanding of their role in essential biological processes, convenient m...

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Autores principales: Sharbati-Tehrani, Soroush, Kutz-Lohroff, Barbara, Bergbauer, Ramona, Scholven, Jutta, Einspanier, Ralf
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2374797/
https://www.ncbi.nlm.nih.gov/pubmed/18400113
http://dx.doi.org/10.1186/1471-2199-9-34
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author Sharbati-Tehrani, Soroush
Kutz-Lohroff, Barbara
Bergbauer, Ramona
Scholven, Jutta
Einspanier, Ralf
author_facet Sharbati-Tehrani, Soroush
Kutz-Lohroff, Barbara
Bergbauer, Ramona
Scholven, Jutta
Einspanier, Ralf
author_sort Sharbati-Tehrani, Soroush
collection PubMed
description BACKGROUND: MicroRNAs (miRNAs) are small endogenous non-coding interfering RNA molecules regarded as major regulators in eukaryotic gene expression. Different methods are employed for miRNA expression profiling. For a better understanding of their role in essential biological processes, convenient methods for differential miRNA expression analysis are required. RESULTS: Here, we present the miR-Q assay as a highly sensitive quantitative reverse transcription PCR (qRT-PCR) for expression analysis of small RNAs such as miRNA molecules. It shows a high dynamic range of 6 to 8 orders of magnitude comprising a sensitivity of up to 0.2 fM miRNA, which corresponds to single copies per cell. There is nearly no cross reaction among closely-related miRNA family members, which points to the high specificity of the assays. Using this approach, we quantified the expression of let-7b in different human cell lines as well as miR-145 and miR-21 expression in porcine intestinal samples. CONCLUSION: miR-Q is a cost-effective and highly specific approach, which neither requires the use of fluorochromic probes, nor Locked Nucleic Acid (LNA)-modified oligonucleotides. Moreover, it provides a remarkable increase in specificity and simplified detection of small RNAs.
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spelling pubmed-23747972008-05-09 miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample Sharbati-Tehrani, Soroush Kutz-Lohroff, Barbara Bergbauer, Ramona Scholven, Jutta Einspanier, Ralf BMC Mol Biol Methodology Article BACKGROUND: MicroRNAs (miRNAs) are small endogenous non-coding interfering RNA molecules regarded as major regulators in eukaryotic gene expression. Different methods are employed for miRNA expression profiling. For a better understanding of their role in essential biological processes, convenient methods for differential miRNA expression analysis are required. RESULTS: Here, we present the miR-Q assay as a highly sensitive quantitative reverse transcription PCR (qRT-PCR) for expression analysis of small RNAs such as miRNA molecules. It shows a high dynamic range of 6 to 8 orders of magnitude comprising a sensitivity of up to 0.2 fM miRNA, which corresponds to single copies per cell. There is nearly no cross reaction among closely-related miRNA family members, which points to the high specificity of the assays. Using this approach, we quantified the expression of let-7b in different human cell lines as well as miR-145 and miR-21 expression in porcine intestinal samples. CONCLUSION: miR-Q is a cost-effective and highly specific approach, which neither requires the use of fluorochromic probes, nor Locked Nucleic Acid (LNA)-modified oligonucleotides. Moreover, it provides a remarkable increase in specificity and simplified detection of small RNAs. BioMed Central 2008-04-10 /pmc/articles/PMC2374797/ /pubmed/18400113 http://dx.doi.org/10.1186/1471-2199-9-34 Text en Copyright © 2008 Sharbati-Tehrani et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methodology Article
Sharbati-Tehrani, Soroush
Kutz-Lohroff, Barbara
Bergbauer, Ramona
Scholven, Jutta
Einspanier, Ralf
miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample
title miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample
title_full miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample
title_fullStr miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample
title_full_unstemmed miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample
title_short miR-Q: a novel quantitative RT-PCR approach for the expression profiling of small RNA molecules such as miRNAs in a complex sample
title_sort mir-q: a novel quantitative rt-pcr approach for the expression profiling of small rna molecules such as mirnas in a complex sample
topic Methodology Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2374797/
https://www.ncbi.nlm.nih.gov/pubmed/18400113
http://dx.doi.org/10.1186/1471-2199-9-34
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