Yamamoto Keisuke, Chiba Mitsuru
Department of Bioscience and Laboratory Medicine, Graduate School of Health Sciences, Hirosaki University, Hirosaki, Aomori 036-8564, Japan.
Research Center for Biomedical Sciences, Hirosaki University, Hirosaki, Aomori 036-8564, Japan.
Biomed Rep. 2024 Jan 30;20(3):51. doi: 10.3892/br.2024.1739. eCollection 2024 Mar.
Serum microRNAs (miRNAs) are considered useful as non-invasive biomarkers for different diseases. However, the optimal method for extracting RNAs from serum is currently unknown. In the present study, several RNA extraction kits were used to examine the optimal kit. RNAs were extracted from the serum of 8-week-old C57BL/6NJcl male mice following the protocol of each RNA extraction kit. The yield of the extracted RNA samples was calculated, and an Agilent Bioanalyzer was used to assess the electrophoretic patterns. An Agilent mouse miRNA microarray was utilized to confirm the expression patterns of the extracted RNA samples. The results revealed significant differences in RNA yields from the miRNeasy Serum/Plasma Advanced kit and mirVana™ PARIS™ RNA and Native Protein Purification Kit compared with almost all other samples. Further, two peaks were determined in the miRNeasy Serum/Plasma Advanced kit using a small RNAs kit of Agilent Bioanalyzer, including one at 20-40 nucleotides (nt) and another at ~40-100 nt, whereas the other reagents had a single peak. This revealed that the extracted RNAs may differ in composition based on the RNA extraction method. Some types of miRNAs were only detected with certain RNA extraction reagents. This suggested that different RNA extraction reagents may cause differences in the types of miRNAs detected. On the other hand, the miRNAs commonly expressed by the three RNA extraction reagents are highly correlated in expression levels.
血清微小RNA（miRNA）被认为是不同疾病的有用非侵入性生物标志物。然而，目前尚不清楚从血清中提取RNA的最佳方法。在本研究中，使用了几种RNA提取试剂盒来检测最佳试剂盒。按照每个RNA提取试剂盒的方案，从8周龄C57BL/6NJcl雄性小鼠的血清中提取RNA。计算提取的RNA样品的产量，并使用安捷伦生物分析仪评估电泳图谱。利用安捷伦小鼠miRNA微阵列确认提取的RNA样品的表达模式。结果显示，与几乎所有其他样品相比，miRNeasy血清/血浆高级试剂盒和mirVana™ PARIS™ RNA和天然蛋白纯化试剂盒的RNA产量存在显著差异。此外，使用安捷伦生物分析仪的小RNA试剂盒在miRNeasy血清/血浆高级试剂盒中确定了两个峰，一个在20 - 40个核苷酸（nt）处，另一个在约40 - 100 nt处，而其他试剂有一个单峰。这表明根据RNA提取方法，提取的RNA在组成上可能有所不同。某些类型的miRNA仅在特定的RNA提取试剂中被检测到。这表明不同的RNA提取试剂可能导致检测到的miRNA类型存在差异。另一方面，三种RNA提取试剂共同表达的miRNA在表达水平上高度相关。
