相关实验视频

Updated: Jul 14, 2025

Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions

Sample Preparation for Mass Spectrometry-based Identification of RNA-binding Regions

Published on: September 28, 2017

通过水友互动色谱学与质谱学相结合,对合成导向核糖核酸进行表征

Bingchuan Wei¹, Jenny Wang¹, Lulu Dai¹

¹Synthetic Molecule Analytical Chemistry, Genentech Inc., 1 DNA Way, South San Francisco, CA 94080, United States of America.

Journal of chromatography. A

|October 8, 2023

概括

这项研究介绍了一种新的水友互动液态染色学 (HILIC) 方法,用于分析单向导核糖核酸 (sgRNA),这对于CRISPR基因组编辑至关重要. HILIC-MS技术有效地表征sgRNA及其聚合物,为寡核酸分析提供了强大的工具.

科学领域:

生物化学生物化学
分析化学分析化学
分子生物学分子生物学

背景情况:

单向导核糖核酸 (sgRNA) 对于CRISPR基因组编辑至关重要.
精确分析sgRNA对于确保基因编辑应用的有效性和安全性至关重要.
现有的分析方法在特征大型寡核酸如sgRNA时可能面临挑战.

研究的目的:

开发和验证用于sgRNA分析的水友交互液体染色学 (HILIC) 方法.
阐明在HILIC中控制sgRNA分离的保留机制.
建立HILIC与质谱学 (HILIC-MS) 结合,作为完整的sgRNA质量分析和杂质分析的强大工具.

主要方法:

开发一种具有压抑静止相表面电荷的HILIC方法,用于sgRNA分析.
研究保留机制,包括结和分离.
直接合HILIC与质谱仪 (MS) 进行完整的质量分析.
使用低温HILIC进行sgRNA聚合物的表征.

主要成果:

通过在HILIC中抑制静止相表面电荷来实现sgRNA的有效配置.
鉴定出结合是大型寡核酸如100-mer sgRNA的主要保留机制.
通过HILIC-MS,可以对sgRNA和杂质进行完整的质量分析,并使用最小的引物.
确定sgRNA聚合物是低温HILIC中后峰的原因.

结论:

开发的HILIC方法提供了sgRNA的有效表征.
HILIC-MS是一种强大的分析工具,用于分析大型寡核酸.
了解保留机制和聚合物形成对于准确的sgRNA分析至关重要.

关键词:

聚合聚合是一种聚合.克里斯普尔是什么意思?克里斯普尔是什么意思?水友互动液体色谱学液体色谱学质谱测量质量谱测量有氧核酸的部分.sgRNARNA 是一个

更多相关视频

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Protocol for the Solid-phase Synthesis of Oligomers of RNA Containing a 2'-O-thiophenylmethyl Modification and Characterization via Circular Dichroism

Published on: July 28, 2017

Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling

Global Identification of Co-Translational Interaction Networks by Selective Ribosome Profiling

Published on: October 7, 2021

相关概念视频

Mass Spectrometry: Complex Analysis

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...

Ribosome Profiling

Ribosome Profiling

Ribosome profiling or ribo-sequencing is a deep sequencing technique that produces a snapshot of active translation in a cell. It selectively sequences the mRNAs protected by ribosomes to get an insight into a cell’s translation landscape at any given point in time.
Applications of ribosome profiling
Ribosome profiling has many applications, including in vivo monitoring of translation inside a particular organ or tissue type and quantifying new protein synthesis levels.
The technique...