Effects of Lyse-It on endonuclease fragmentation, function and activity
dc.contributor.author | Santaus, Tonya M. | |
dc.contributor.author | Zhang, Fan | |
dc.contributor.author | Li, Shan | |
dc.contributor.author | Stine, O. Colin | |
dc.contributor.author | Geddes, Chris D. | |
dc.date.accessioned | 2019-11-04T17:57:41Z | |
dc.date.available | 2019-11-04T17:57:41Z | |
dc.date.issued | 2019-09-30 | |
dc.description.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. | en_US |
dc.description.sponsorship | This work is supported by the National Institutes of Health UMBC Graduate Training Chemistry Biology Interface Fellowship (T32GM066706-15). We note that the University of Maryland, Baltimore County (UMBC) has filed and has had issued several patents related to microwave-based lysing and DNA fragmentation. We note that those patents are licensed to Lyse-It LLC, a Maryland-based biotechnology company to which Dr. Geddes currently owns stock. Lyse-It LLC was not involved in the study design of the material within this paper, nor do any of the author receive any salaries or consultancies from Lyse-It LLC. This does not alter our adherence to PLOS ONE policies on sharing data and materials | en_US |
dc.description.uri | https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0223008 | en_US |
dc.format.extent | 21 pages | en_US |
dc.genre | journal articles | en_US |
dc.identifier | doi:10.13016/m2ijeu-g4fa | |
dc.identifier.citation | 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 | en_US |
dc.identifier.uri | https://doi.org/10.1371/journal.pone.0223008 | |
dc.identifier.uri | http://hdl.handle.net/11603/16031 | |
dc.language.iso | en_US | en_US |
dc.publisher | PLOS | en_US |
dc.relation.isAvailableAt | The University of Maryland, Baltimore County (UMBC) | |
dc.relation.ispartof | UMBC Chemistry & Biochemistry Department Collection | |
dc.relation.ispartof | UMBC Institute of Fluorescence (IoF) | |
dc.relation.ispartof | UMBC Faculty Collection | |
dc.rights | This item is likely protected under Title 17 of the U.S. Copyright Law. Unless on a Creative Commons license, for uses protected by Copyright Law, contact the copyright holder or the author. | |
dc.rights | Attribution 4.0 International (CC BY 4.0) | * |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | * |
dc.subject | Nucleases | en_US |
dc.subject | Ribonucleases | en_US |
dc.subject | Microwave radiation | en_US |
dc.subject | Deoxyribonucleases | en_US |
dc.subject | SDS polyacrylamide gel electrophoresis | en_US |
dc.subject | Oxygen | en_US |
dc.subject | Reactive oxygen species | en_US |
dc.subject | Vibrio cholerae | en_US |
dc.title | Effects of Lyse-It on endonuclease fragmentation, function and activity | en_US |
dc.type | Text | en_US |
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