聚乙二醇介导的外泌体分离：外泌体 RNA 分析的一种方法。
Polyethylene Glycol -Mediated Exosome Isolation: A Method for Exosomal RNA Analysis.
出版信息
Iran Biomed J. 2024 Mar 1;28(2&3):132-9. doi: 10.61186/ibj.4129.
BACKGROUND
: Exosomal RNAs (ExoRNAs) offer valuable insights into their cellular origin. ExoRNA studies were faced with challenges in obtaining sufficient amounts of high-quality RNA. Herein, we aimed to compare three traditional exosome isolation methods to introduce an appropriate strategy to extract RNA from cancer-derived exosomes for further RNA analysis.
METHODS
Exosomes were isolated through ultracentrifugation, precipitation kit, and size exclusion column chromatography, and then characterized by dynamic light scattering and transmission electron microscopy, followed by extracting total RNA. The quality and quantity of the extracted RNAs were assessed by a NanoDrop and 2.5% agarose gel electrophoresis.
RESULTS
Extracted exosomes displayed a similar range of size and morphology. We found that polyethylene glycol-precipitation method resulted in a higher RNA yield with a 260/280 ratio of 1.9. The obtained exoRNA appeared as a smear in the agarose gel, indicative of small exoRNAs.
CONCLUSION
We provide researchers a suitable approach to isolate exosomes based on yield and purity of exoRNA.
背景
外泌体中的 RNA（ExoRNAs）可以为其细胞起源提供有价值的见解。外泌体 RNA 研究在获得足够数量的高质量 RNA 方面面临挑战。在此，我们旨在比较三种传统的外泌体分离方法，以引入一种从癌症来源的外泌体中提取 RNA 的合适策略，用于进一步的 RNA 分析。
方法
通过超速离心、沉淀试剂盒和大小排阻柱色谱法分离外泌体，然后通过动态光散射和透射电子显微镜进行表征，接着提取总 RNA。通过 NanoDrop 和 2.5%琼脂糖凝胶电泳评估提取的 RNA 的质量和数量。
结果
提取的外泌体显示出相似的大小和形态范围。我们发现聚乙二醇沉淀法可获得更高的 RNA 产量，260/280 比值为 1.9。获得的外泌体 RNA 在琼脂糖凝胶上呈现出弥散状，表明是小外泌体 RNA。
结论
我们为研究人员提供了一种基于外泌体 RNA 产量和纯度的合适的外泌体分离方法。
Zotero 插件