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Evaluation of a novel approach for the measurement of RNA quality

BACKGROUND: Microarray data interpretation can be affected by sample RNA integrity. The ScreenTape Degradation Value (SDV) is a novel RNA integrity metric specific to the ScreenTape(® )platform (Lab901). To characterise the performance of the ScreenTape(® )platform for RNA analysis and determine the...

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Autores principales: Wilkes, Timothy M, Devonshire, Alison S, Ellison, Stephen LR, Foy, Carole A
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2907863/
https://www.ncbi.nlm.nih.gov/pubmed/20359344
http://dx.doi.org/10.1186/1756-0500-3-89
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author Wilkes, Timothy M
Devonshire, Alison S
Ellison, Stephen LR
Foy, Carole A
author_facet Wilkes, Timothy M
Devonshire, Alison S
Ellison, Stephen LR
Foy, Carole A
author_sort Wilkes, Timothy M
collection PubMed
description BACKGROUND: Microarray data interpretation can be affected by sample RNA integrity. The ScreenTape Degradation Value (SDV) is a novel RNA integrity metric specific to the ScreenTape(® )platform (Lab901). To characterise the performance of the ScreenTape(® )platform for RNA analysis and determine the robustness of the SDV metric, a panel of intentionally degraded RNA samples was prepared. These samples were used to evaluate the ScreenTape(® )platform against an alternative approach for measuring RNA integrity (Agilent Bioanalyzer RIN value). The samples were also subjected to microarray analysis and the resulting data correlated to the RNA integrity metrics. FINDINGS: Measurement of SDV for a panel of intentionally degraded RNA samples ranged from 0 for intact RNA to 37 for degraded RNA, with corresponding RIN values ranging from 10 to 4 for the same set of samples. SDV and RIN scales both demonstrated comparable discrimination between differently treated samples (RIN 10 to 7, SDV 0 to 15), with the SDV exhibiting better discrimination at higher degradation levels. Increasing SDV values correlated with a decrease in microarray sample labelling efficiency and an increase in numbers of differentially expressed genes. CONCLUSIONS: The ScreenTape(® )platform is comparable to the Bioanalyzer platform in terms of reproducibility and discrimination between different levels of RNA degradation. The robust nature of the SDV metric qualifies it as an alternative metric for RNA sample quality control, and a useful predictor of downstream microarray performance.
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spelling pubmed-29078632010-07-22 Evaluation of a novel approach for the measurement of RNA quality Wilkes, Timothy M Devonshire, Alison S Ellison, Stephen LR Foy, Carole A BMC Res Notes Short Report BACKGROUND: Microarray data interpretation can be affected by sample RNA integrity. The ScreenTape Degradation Value (SDV) is a novel RNA integrity metric specific to the ScreenTape(® )platform (Lab901). To characterise the performance of the ScreenTape(® )platform for RNA analysis and determine the robustness of the SDV metric, a panel of intentionally degraded RNA samples was prepared. These samples were used to evaluate the ScreenTape(® )platform against an alternative approach for measuring RNA integrity (Agilent Bioanalyzer RIN value). The samples were also subjected to microarray analysis and the resulting data correlated to the RNA integrity metrics. FINDINGS: Measurement of SDV for a panel of intentionally degraded RNA samples ranged from 0 for intact RNA to 37 for degraded RNA, with corresponding RIN values ranging from 10 to 4 for the same set of samples. SDV and RIN scales both demonstrated comparable discrimination between differently treated samples (RIN 10 to 7, SDV 0 to 15), with the SDV exhibiting better discrimination at higher degradation levels. Increasing SDV values correlated with a decrease in microarray sample labelling efficiency and an increase in numbers of differentially expressed genes. CONCLUSIONS: The ScreenTape(® )platform is comparable to the Bioanalyzer platform in terms of reproducibility and discrimination between different levels of RNA degradation. The robust nature of the SDV metric qualifies it as an alternative metric for RNA sample quality control, and a useful predictor of downstream microarray performance. BioMed Central 2010-04-01 /pmc/articles/PMC2907863/ /pubmed/20359344 http://dx.doi.org/10.1186/1756-0500-3-89 Text en Copyright ©2010 Wilkes 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 Short Report
Wilkes, Timothy M
Devonshire, Alison S
Ellison, Stephen LR
Foy, Carole A
Evaluation of a novel approach for the measurement of RNA quality
title Evaluation of a novel approach for the measurement of RNA quality
title_full Evaluation of a novel approach for the measurement of RNA quality
title_fullStr Evaluation of a novel approach for the measurement of RNA quality
title_full_unstemmed Evaluation of a novel approach for the measurement of RNA quality
title_short Evaluation of a novel approach for the measurement of RNA quality
title_sort evaluation of a novel approach for the measurement of rna quality
topic Short Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2907863/
https://www.ncbi.nlm.nih.gov/pubmed/20359344
http://dx.doi.org/10.1186/1756-0500-3-89
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