常用氧气清除系统中核酸酶活性的缺失。
Absence of nuclease activity in commonly used oxygen-scavenging systems.
作者信息
Gahlon Hailey L, Poudevigne-Durance Paul, Rueda David
机构信息
Molecular Virology, Department of Medicine, Imperial College London, Du Cane Road, London, W12 0NN, UK.
Single Molecule Imaging Group, MRC London Institute of Medical Sciences, Du Cane Road, London, W12 0NN, UK.
出版信息
BMC Res Notes. 2017 Nov 21;10(1):606. doi: 10.1186/s13104-017-2929-6.
OBJECTIVE
Oxygen scavenging systems are routinely used during single-molecule imaging experiments to improve fluorescent dye stability. Previous work has shown nuclease contamination in the commonly used oxygen scavenging systems. This study evaluates the potential for nuclease contamination in these oxygen scavenging systems.
RESULTS
Linear and plasmid DNA was incubated with two different oxygen scavenging systems (1) protocatechuic acid (PCA)-protocatechuate-3,4-dioxygenase (PCD) and (2) glucose-coupled glucose oxidase/catalase (GODCAT). No nucleic acid degradation was observed on single and double-stranded linear DNA and plasmid DNA, indicating the absence of nuclease contamination in these oxygen scavenging systems.
目的
在单分子成像实验中，通常会使用氧气清除系统来提高荧光染料的稳定性。先前的研究表明，常用的氧气清除系统中存在核酸酶污染。本研究评估了这些氧气清除系统中核酸酶污染的可能性。
结果
将线性和质粒DNA与两种不同的氧气清除系统（1）原儿茶酸（PCA）-原儿茶酸-3,4-双加氧酶（PCD）和（2）葡萄糖偶联葡萄糖氧化酶/过氧化氢酶（GODCAT）一起孵育。在单链和双链线性DNA以及质粒DNA上均未观察到核酸降解，表明这些氧气清除系统中不存在核酸酶污染。
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