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

Effects of Lyse-It on endonuclease fragmentation, function and activity

Files

Original bundle

License bundle

Collections