Banner Sun Health Research Institute, Sun City, AZ 85351, USA.
Int J Mol Sci. 2023 Sep 7;24(18):13795. doi: 10.3390/ijms241813795.
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 r 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 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.
RNA 完整性的测定是基因表达研究的一个关键质量评估工具,实验的成功高度依赖于样本质量。自 1999 年推出以来,科学界的金标准一直是安捷伦 2100 Bioanalyzer 的 RNA 完整性号码(RIN),它使用 1-10 的数值系统,1 表示降解最严重,10 表示最完整。2015 年,安捷伦推出了 4200 TapeStation 的 RIN 等效值,并报告了 r 值为 0.936 的强相关性和中位数误差<±0.4 RIN 单位。为了评估这一说法,我们比较了安捷伦 4200 TapeStation 的 RIN 等效值(RINe)和 DV200 与安捷伦 2100 Bioanalyzer 的 RIN 在 183 个平行 RNA 样本中的相关性。在我们的研究中,使用总共 183 个人死后大脑样本的 RNA,我们发现 RIN 和 RINe 值相关性较弱,r 值为 0.393,平均差异为 3.2 RIN 单位。DV200 与 RIN(r 为 0.182)和 RINe(r 为 0.347)也只有微弱的相关性。最后,当对这两个指标都使用 6.5 的截止值时,我们发现 95.6%的样本通过了 RIN,而只有 23.5%的样本通过了 RINe。我们的结果表明,尽管 RIN(Bioanalyzer)和 RINe(TapeStation)使用相同的 1-10 值系统,但它们不应互换使用,并且应独立计算截止值。