Measuring Up: A Comparison of TapeStation 4200 and Bioanalyzer 2100 as Measurement Tools for RNA Quality in Postmortem Human Brain Samples

Author:

Walker Jessica E.1,Oliver Javon C.1,Stewart Analisa M.1,Beh Suet Theng1,Arce Richard A.1,Glass Michael J.1,Vargas Daisy E.1,Qiji Sanaria H.1,Intorcia Anthony J.1,Borja Claryssa I.1,Cline Madison P.1,Hemmingsen Spencer J.1,Krupp Addison N.1,McHattie Rylee D.1,Mariner Monica R.1,Lorenzini Ileana1,Aslam Sidra1,Tremblay Cecilia1,Beach Thomas G.1,Serrano Geidy E.1

Affiliation:

1. Banner Sun Health Research Institute, Sun City, AZ 85351, USA

Abstract

The determination of RNA integrity is a critical quality assessment tool for gene expression studies where the experiment’s success is highly dependent on the sample quality. Since its introduction in 1999, the gold standard in the scientific community has been the Agilent 2100 Bioanalyzer’s RNA integrity number (RIN), which uses a 1–10 value system, from 1 being the most degraded, to 10 being the most intact. In 2015, Agilent launched 4200 TapeStation’s RIN equivalent, and reported a strong correlation of r2 of 0.936 and a median error < ±0.4 RIN units. To evaluate this claim, we compared the Agilent 4200 TapeStation’s RIN equivalent (RINe) and DV200 to the Agilent 2100 Bioanalyzer’s RIN for 183 parallel RNA samples. In our study, using RNA from a total of 183 human postmortem brain samples, we found that the RIN and RINe values only weakly correlate, with an r2 of 0.393 and an average difference of 3.2 RIN units. DV200 also only weakly correlated with RIN (r2 of 0.182) and RINe (r2 of 0.347). Finally, when applying a cut-off value of 6.5 for both metrics, we found that 95.6% of samples passed with RIN, while only 23.5% passed with RINe. Our results suggest that even though RIN (Bioanalyzer) and RINe (TapeStation) use the same 1–10 value system, they should not be used interchangeably, and cut-off values should be calculated independently.

Funder

the National Institute of Neurological Disorders and Stroke

the National Institute on Aging

the Arizona Department of Health Services

the Arizona Biomedical Research Commission

the Michael J. Fox Foundation for Parkinson’s Research

Publisher

MDPI AG

Subject

Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis

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