单分子并行分析用于快速探索序列空间
Carolien Bastiaanssen1, Ivo Severins1,2,3, John van Noort4
1Department of BioNanoScience, Kavli Institute of Nanoscience, Delft University of Technology, Van der Maasweg 9, Delft, the Netherlands.
Nature protocols
|June 4, 2025
概括
我们开发了SPARXS,这是一个单分子分析的高通量平台,可以快速探索DNA和RNA序列空间. 这种方法同时分析数百万个分子,揭示了依赖序列的生物分子动力学.
科学领域:
- 生物物理学的生物物理.
- 分子生物学分子生物学
- 基因组学就是基因组学.
背景情况:
- 单分子光技术揭示了生物分子结构和动态.
- 由于时间和成本的限制,目前的方法仅限于少数序列.
- 了解序列影响需要对众多序列进行高通量分析.
研究的目的:
- 开发一个高通量平台,在单个分子层面探索序列空间.
- 为了使数以百万计的分子在数千个序列中同时进行分析.
- 研究对分子过程的序列效应,例如霍莱德连接动力学.
主要方法:
- 开发了SPARXS (用于快速探索序列空间的单分子并行分析).
- 结合单分子光显微镜与下一代高通量测序.
- 详细的样本设计,单分子测量,测序,数据集成和分析使用Papylio包的详细协议.
主要成果:
- 在单个分子层面上,SPARXS可以同时对数百万个分子进行分析.
- 该平台涵盖数千个不同的序列,提供定量见解.
- 在研究对霍莱德连接运动的序列效应中得到了应用.
结论:
- SPARXS显著提高了研究生物分子中的序列结构功能关系的能力.
- 该平台提供了一种快速,具有成本效益的方法来探索庞大的序列空间.
- 提供了对分子动力学和相互作用的序列影响的详细定量图片.
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