Whole blood is one of the most widely utilized human samples in biological research and is useful for analyzing the mechanisms of diverse bio-molecular phenomena. However, owing to its fluidic properties, whole blood is relatively unstable in the frozen state compared to other samples. Because RNA is structurally unstable, sample damage can severely affect RNA quality, thereby reducing its usability. This study aimed to assess the quality of RNA which was prepared from blood stored at different temperatures and times prior to freezing, as well as the effect of long-term freezing. The quality of the RNA derived from different blood samples was assessed by measuring the RNA integrity number (RIN) and RNA sequencing to identify differentially expressed genes (DEGs, |fold-change (FC)|> 1.5, p < 0.05, false discovery rate (FDR) < 0.05) between the differently prepared RNA samples. We found that improper sample handling critically influenced both RNA quality and gene expression patterns. By suggesting the consequences, this study emphasizes the importance of sample management to obtain reliable downstream application outcomes.

全血是生物学研究中使用最广泛的人类样品之一，可用于分析多种生物分子现象的机制。但是，由于其流体特性，与其他样品相比，全血在冷冻状态下相对不稳定。由于RNA在结构上不稳定，因此样品损坏会严重影响RNA的质量，从而降低其可用性。这项研究旨在评估从冷冻前在不同温度和时间存储的血液中制备的RNA的质量，以及长期冷冻的效果。通过测量RNA完整性数（RIN）和RNA测序以鉴定差异表达的基因（DEGs，|倍数变化（FC）|> 1.5，p <0.05，错误发现率），评估了来自不同血样的RNA的质量。 （FDR）<0。05）不同制备的RNA样品之间。我们发现不正确的样品处理会严重影响RNA质量和基因表达模式。通过暗示后果，本研究强调了样品管理对获得可靠的下游应用结果的重要性。