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DV200 Index for Assessing RNA Integrity in Next-Generation Sequencing
Joint Authors
Matsubara, Takehiro
Soh, Junichi
Morita, Mizuki
Uwabo, Takahiro
Tomida, Shuta
Fujiwara, Toshiyoshi
Toyooka, Shinichi
Hirasawa, Akira
Kanazawa, Susumu
Source
Issue
Vol. 2020, Issue 2020 (31 Dec. 2020), pp.1-6, 6 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2020-02-27
Country of Publication
Egypt
No. of Pages
6
Main Subjects
Abstract EN
Poor quality of biological samples will result in an inaccurate analysis of next-generation sequencing (NGS).
Therefore, methods to accurately evaluate sample integrity are needed.
Among methods for evaluating RNA quality, the RNA integrity number equivalent (RINe) is widely used, whereas the DV200, which evaluates the percentage of fragments of >200 nucleotides, is also used as a quality assessment standard.
In this study, we compared the RINe and DV200 RNA quality indexes to determine the most suitable RNA index for the NGS analysis.
Seventy-one RNA samples were extracted from formalin-fixed paraffin-embedded tissue samples (n=30), fresh-frozen samples (n=25), or cell lines (n=16).
After assessing RNA quality using the RINe and DV200, we prepared two kinds of stranded mRNA sequencing libraries.
Finally, we calculated the correlation between each RNA quality index and the amount of library product (1st PCR product per input RNA).
The DV200 measure showed stronger correlation with the amount of library product than the RINe (R2=0.8208 for the DV200 versus 0.6927 for the RINe).
Receiver operating characteristic curve analyses revealed that the DV200 was the better marker for predicting efficient library production than the RINe using a threshold of >10 ng/ng for the amount of the 1st PCR product per input RNA (cutoff value for the RINe and DV200, 2.3 and 66.1%; area under the curve, 0.99 and 0.91; sensitivity, 82% and 92%; and specificity, 93% and 100%, respectively).
Our results indicate that NGS libraries prepared using RNA samples with the DV200 value>66.1% exhibit greater sensitivity and specificity than those prepared with the RINe values>2.3.
These findings suggest that the DV200 is superior to the RINe, especially for low-quality RNA, because it is a more consistent assessment of the amount of the 1st NGS library product per input.
American Psychological Association (APA)
Matsubara, Takehiro& Soh, Junichi& Morita, Mizuki& Uwabo, Takahiro& Tomida, Shuta& Fujiwara, Toshiyoshi…[et al.]. 2020. DV200 Index for Assessing RNA Integrity in Next-Generation Sequencing. BioMed Research International،Vol. 2020, no. 2020, pp.1-6.
https://search.emarefa.net/detail/BIM-1138078
Modern Language Association (MLA)
Matsubara, Takehiro…[et al.]. DV200 Index for Assessing RNA Integrity in Next-Generation Sequencing. BioMed Research International No. 2020 (2020), pp.1-6.
https://search.emarefa.net/detail/BIM-1138078
American Medical Association (AMA)
Matsubara, Takehiro& Soh, Junichi& Morita, Mizuki& Uwabo, Takahiro& Tomida, Shuta& Fujiwara, Toshiyoshi…[et al.]. DV200 Index for Assessing RNA Integrity in Next-Generation Sequencing. BioMed Research International. 2020. Vol. 2020, no. 2020, pp.1-6.
https://search.emarefa.net/detail/BIM-1138078
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1138078