酸性无凝胶毛细管区带电泳用于微DNA和RNA分析

IF 5.6 1区化学 Q1 CHEMISTRY, ANALYTICAL

Talanta Pub Date : 2025-07-21 DOI:10.1016/j.talanta.2025.128610

Jakob Haglöf , Erik Bivehed , Cari Sänger-van de Griend

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引用次数: 0

摘要

短的DNA和RNA序列，如治疗性反义寡核苷酸（ASO），微DNA （miDNA）和microRNA （miRNA）主要使用液相色谱技术或毛细管凝胶电泳进行分析。目前对改进这些核苷酸分析的需求促使人们重新考虑旧的真理，例如需要凝胶基毛细管电泳进行DNA和RNA分析。本研究表明，使用酸性背景电解质的毛细管区带电泳可用于分离ASO-， miDNA-和mirna大小的核苷酸。利用不同的核碱基pKa作为基础，成功分离出大小相同但序列不同的不同核苷酸，效率高，分析时间短。此外，腺苷和胞苷与鸟苷和胸腺嘧啶核碱基含量的比值AC/GT被证明是评估单链核苷酸迁移和分离的有效参数。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

Gel-free capillary zone electrophoresis at acidic pH for micro DNA and RNA analysis

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Gel-free capillary zone electrophoresis at acidic pH for micro DNA and RNA analysis

Short DNA and RNA sequences like therapeutic anti-sense oligonucleotides (ASO), microDNA (miDNA) and microRNA (miRNA) are predominantly analyzed using liquid chromatographic techniques or capillary gel electrophoresis. The current demand for improved analysis of these nucleotides gives incentives to reconsider old truths, such as the need for gel-based capillary electrophoresis for DNA and RNA analysis.

The present study shows the applicability of capillary zone electrophoresis using acidic background electrolytes for the separation of ASO-, miDNA- and miRNA-sized nucleotides. Using the different pKa of the nucleobases as a foundation, different nucleotides with same size, but varying sequence, has been successfully separated with high efficiency and short analysis time. Furthermore, the ratio of adenosine and cytidine to guanosine and thymidine nucleobase content, AC/GT, is shown as a valid parameter to assess single-stranded nucleotide migration and separation.

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来源期刊

Talanta 化学-分析化学

CiteScore

12.30

自引率

4.90%

发文量

861

审稿时长

29 days

期刊介绍： Talanta provides a forum for the publication of original research papers, short communications, and critical reviews in all branches of pure and applied analytical chemistry. Papers are evaluated based on established guidelines, including the fundamental nature of the study, scientific novelty, substantial improvement or advantage over existing technology or methods, and demonstrated analytical applicability. Original research papers on fundamental studies, and on novel sensor and instrumentation developments, are encouraged. Novel or improved applications in areas such as clinical and biological chemistry, environmental analysis, geochemistry, materials science and engineering, and analytical platforms for omics development are welcome. Analytical performance of methods should be determined, including interference and matrix effects, and methods should be validated by comparison with a standard method, or analysis of a certified reference material. Simple spiking recoveries may not be sufficient. The developed method should especially comprise information on selectivity, sensitivity, detection limits, accuracy, and reliability. However, applying official validation or robustness studies to a routine method or technique does not necessarily constitute novelty. Proper statistical treatment of the data should be provided. Relevant literature should be cited, including related publications by the authors, and authors should discuss how their proposed methodology compares with previously reported methods.