Abstract
ObjectiveOxygen 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.ResultsLinear 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.
Highlights
Previous work has suggested that contaminant nuclease enzymes could be affecting single-molecule studies [1]
Using size-exclusion chromatography, this study reported the presence of nuclease contamination in the 40- and 100 kDa elution fractions of routinely used oxygen scavenging system (OSS) system with protocatechuic acid (PCA)-protocatechuate-3,4-dioxygenase (PCD), with the 40 kDa fraction exhibiting the highest nuclease activity
Linear, single- and double-stranded, and plasmid DNA substrates were tested for nuclease contamination in oxygen scavenging systems that are routinely used during single-molecule imaging
Summary
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).
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