The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.
Towards standardization of RNA quality assessment using user-independent classifiers of microcapillary electrophoresis traces
Open accessAbstract
While it is universally accepted that intact RNA constitutes the best representation of the steady-state of transcription, there is no gold standard to define RNA quality prior to gene expression analysis. In this report, we evaluated the reliability of conventional methods for RNA quality assessment including UV spectroscopy and 28S:18S area ratios, and demonstrated their inconsistency. We then used two new freely available classifiers, the Degradometer and RIN systems, to produce user-independent RNA quality metrics, based on analysis of microcapillary electrophoresis traces. Both provided highly informative and valuable data and the results were found highly correlated, while the RIN system gave more reliable data. The relevance of the RNA quality metrics for assessment of gene expression differences was tested by Q-PCR, revealing a significant decline of the relative expression of genes in RNA samples of disparate quality, while samples of similar, even poor integrity were found highly comparable. We discuss the consequences of these observations to minimize artifactual detection of false positive and negative differential expression due to RNA integrity differences, and propose a scheme for the development of a standard operational procedure, with optional registration of RNA integrity metrics in public repositories of gene expression data.
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Cites
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Cited by
The RIN: an RNA integrity number for assigning integrity values to RNA measurements
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“The calculation of RIN value is largely based on ribosomal RNA separation although this measure has been shown to be inconsistent (Imbeaud et al. 2005; Schroeder et al. 2006; Sherwood et al. 2011).”
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Bias in recent miRBase annotations potentially associated with RNA quality issues
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Metadata
- Authors
- Imbeaud, S.
- Venue
- Nucleic Acids Research
- Publisher
- Oxford University Press (OUP)
- Published
- 2005-03-23
- Volume
- 33
- Issue
- 6
- Pages
- e56-e56
- ISSN
- 0305-1048 (ISSN-L)
- 1362-4962
- OpenAlex
- W2096976625
- Cited by
- 528
- References
- 2
- FWCI
- 10.14
- Type
- journal article
- Language
- en
- Open access
- gold
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