Isolation of short RNAs with homogenous 3’-ends using quaternary-amine anion exchange chromatography

IF 4.7 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-05-17 DOI:10.1093/biomethods/bpae033

Zixian Li, Mia Bilic, Bhushan Nagar

引用次数: 0

Abstract

Visualizing RNA-protein interactions through structural approaches requires the use of RNA molecules purified to homogeneity. We describe here a simple and effective method, free of acrylamide contamination and without using UV radiation, to separate in vitro synthesized, heterogeneous RNA transcripts (up to ∼15 nucleotides) at single nucleotide resolution by quaternary-amine anion exchange chromatography. The quality of short RNAs isolated through this method is validated by gel electrophoresis, mass spectrometry, and crystallization with a protein binding partner.

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利用季胺阴离子交换色谱法分离具有同源 3'- 末端的短 RNA

通过结构方法观察 RNA 蛋白相互作用需要使用纯化到均一的 RNA 分子。我们在此介绍一种简单有效的方法，该方法无丙烯酰胺污染，无需紫外线辐射，可通过季胺阴离子交换色谱法以单核苷酸分辨率分离体外合成的异质 RNA 转录本（最多 15 个核苷酸）。通过凝胶电泳、质谱分析和与蛋白质结合的结晶验证了用这种方法分离出的短 RNA 的质量。

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来源期刊

ACS Applied Bio Materials Chemistry-Chemistry (all)

CiteScore

9.40

自引率

2.10%

发文量

464

期刊介绍： ACS Applied Bio Materials is an interdisciplinary journal publishing original research covering all aspects of biomaterials and biointerfaces including and beyond the traditional biosensing, biomedical and therapeutic applications. The journal is devoted to reports of new and original experimental and theoretical research of an applied nature that integrates knowledge in the areas of materials, engineering, physics, bioscience, and chemistry into important bio applications. The journal is specifically interested in work that addresses the relationship between structure and function and assesses the stability and degradation of materials under relevant environmental and biological conditions.