Liu Zhiyu, Ouyang Tinglan, Yang Yuwei, Sheng Yuqi, Shi Huajuan, Liu Quanjun, Bai Yunfei, Ge Qinyu
State Key Laboratory of Digital Medical Engineering, Southeast University, Nanjing 211189, China.
Genes (Basel). 2024 Apr 17;15(4):502. doi: 10.3390/genes15040502.
In gene quantification and expression analysis, issues with sample selection and processing can be serious, as they can easily introduce irrelevant variables and lead to ambiguous results. This study aims to investigate the extent and mechanism of the impact of sample selection and processing on ribonucleic acid (RNA) sequencing. RNA from PBMCs and blood samples was investigated in this study. The integrity of this RNA was measured under different storage times. All the samples underwent high-throughput sequencing for comprehensive evaluation. The differentially expressed genes and their potential functions were analyzed after the samples were placed at room temperature for 0h, 4h and 8h, and different feature changes in these samples were also revealed. The sequencing results showed that the differences in gene expression were higher with an increased storage time, while the total number of genes detected did not change significantly. There were five genes showing gradient patterns over different storage times, all of which were protein-coding genes that had not been mentioned in previous studies. The effect of different storage times on seemingly the same samples was analyzed in this present study. This research, therefore, provides a theoretical basis for the long-term consideration of whether sample processing should be adequately addressed.
在基因定量和表达分析中，样本选择和处理问题可能很严重，因为它们很容易引入无关变量并导致结果模糊。本研究旨在调查样本选择和处理对核糖核酸（RNA）测序的影响程度和机制。本研究对来自外周血单核细胞（PBMC）和血液样本的RNA进行了研究。在不同储存时间下测量了该RNA的完整性。所有样本都进行了高通量测序以进行全面评估。在样本在室温下放置0小时、4小时和8小时后，分析了差异表达基因及其潜在功能，同时也揭示了这些样本中的不同特征变化。测序结果表明，随着储存时间的增加，基因表达差异更大，而检测到的基因总数没有显著变化。有五个基因在不同储存时间呈现梯度模式，所有这些都是先前研究中未提及的蛋白质编码基因。本研究分析了不同储存时间对看似相同样本的影响。因此，本研究为长期考虑样本处理是否应得到充分解决提供了理论依据。
