Department of Chemical Engineering, Carnegie Mellon University, Pittsburgh, Pennsylvania 15213, United States.
Biomacromolecules. 2024 Aug 12;25(8):4891-4897. doi: 10.1021/acs.biomac.4c00332. Epub 2024 Jul 17.
We demonstrate that the attachment of 30-170 bp dsDNA oligomers to ssDNA viral genomes gives a significant additional mobility shift in micelle-tagging electrophoresis (MTE). In MTE, a modified peptide nucleic acid amphiphile is attached to the viral genome to bind drag-inducing micelles present in capillary electrophoresis running buffers. Further attachment of 30-170 bp dsDNA oligomers drastically shifts the mobility of the 5.1 kB ssDNA genome of mouse minute virus (MMV), providing a new mechanism to improve resolution in CE-based analysis of kilobase nucleic acids. A model based on biased-reptation electrophoresis, end-labeled free-solution electrophoresis, and Ferguson gel-filtration theory is presented to describe the observed mobility shifts.
我们证明,将 30-170bp 的双链 DNA 寡核苷酸与单链 DNA 病毒基因组结合,在胶束标记电泳 (MTE) 中会产生显著的额外迁移率变化。在 MTE 中,一种修饰的肽核酸两亲分子与病毒基因组结合,以结合毛细管电泳运行缓冲液中存在的诱导拖曳的胶束。进一步结合 30-170bp 的双链 DNA 寡核苷酸会极大地改变 5.1kb 的单链 DNA 基因组的迁移率,为基于 CE 的千碱基核酸分析中提高分辨率提供了一种新机制。提出了一个基于有偏的缠结电泳、末端标记的自由溶液电泳和 Ferguson 凝胶过滤理论的模型,以描述观察到的迁移率变化。