Effects of Lyse-It on endonuclease fragmentation, function and activity
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Author/Creator ORCID
Date
2019-09-30
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Citation of Original Publication
Santaus TM, Zhang F, Li S, Stine OC, Geddes CD (2019) Effects of Lyse-It on endonuclease fragmentation, function and activity. PLoS ONE 14(9): e0223008. https://doi.org/ 10.1371/journal.pone.0223008
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Attribution 4.0 International (CC BY 4.0)
Attribution 4.0 International (CC BY 4.0)
Abstract
Nucleases are enzymes that can degrade genomic DNA and RNA that decrease the accuracy of quantitative measures of those nucleic acids. Here, we study conventional heating, standard microwave irradiation, and Lyse-It, a microwave-based lysing technology, for the potential to fragment and inactivate DNA and RNA endonucleases. Lyse-It employs the use of highly focused microwave irradiation to the sample ultimately fragmenting and inactivating RNase A, RNase B, and DNase I. Nuclease size and fragmentation were determined visually and quantitatively by SDS polyacrylamide gel electrophoresis and the mini-gel Agilent 2100 Bioanalyzer system, with a weighted size calculated to depict the wide range of nuclease fragmentation. Enzyme activity assays were conducted, and the rates were calculated to determine the effect of various lysing conditions on each of the nucleases. The results shown in this paper clearly demonstrate that Lyse-It is a rapid and highly efficient way to degrade and inactivate nucleases so that nucleic acids can be retained for down-stream detection.