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对比评估:Tapestation 4200和Bioanalyzer 2100作为死后人类脑样本RNA质量测量工具的比较

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

作者信息

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

出版信息

medRxiv. 2023 Jul 6:2023.07.03.23291969. doi: 10.1101/2023.07.03.23291969.

Abstract

Determining RNA integrity is a critical quality assessment tool for gene expression studies where the experiment's success is highly dependent on 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 with 1 being the most degraded to 10 being the most intact. In 2015, Agilent launched the 4200 Tapestation's RIN equivalent and reported a strong correlation of r of 0.936 and 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 r of 0.393 and an average difference of 3.2 RIN units. DV200 also only weakly correlated with RIN (r of 0.182) and RINe (r 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.

摘要

确定RNA完整性是基因表达研究中一项关键的质量评估工具,该实验的成功高度依赖于样本质量。自1999年引入以来,科学界的金标准一直是安捷伦2100生物分析仪的RNA完整性数值(RIN),它采用1至10的数值系统,1表示降解程度最高,10表示最完整。2015年,安捷伦推出了4200 TapeStation的等效RIN,并报告其与安捷伦2100生物分析仪的RIN有很强的相关性,r值为0.936,中位数误差<±0.4 RIN单位。为了评估这一说法,我们将183个平行RNA样本的安捷伦4200 TapeStation的等效RIN(RINe)和DV200与安捷伦2100生物分析仪的RIN进行了比较。在我们的研究中,使用总共183个人类死后大脑样本的RNA,我们发现RIN和RINe值仅呈弱相关,r值为0.393,平均差异为3.2个RIN单位。DV200与RIN(r值为0.182)和RINe(r值为0.347)也仅呈弱相关。最后,当对这两个指标都应用6.5的临界值时,我们发现95.6%的样本通过了RIN检测,而只有23.5%的样本通过了RINe检测。我们的结果表明,尽管RIN(生物分析仪)和RINe(TapeStation)使用相同的1至10数值系统,但它们不应互换使用,临界值应独立计算。

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