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Effects of Lyse-It on endonuclease fragmentation, function and activity
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Effects of Lyse-It on endonuclease fragmentation, function and activity
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Journal Article
Effects of Lyse-It on endonuclease fragmentation, function and activity
2019
Overview
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.
Publisher
Public Library of Science,Public Library of Science (PLoS)
Subject
/ Biology
/ Biomedical laboratory equipment
/ Deoxyribonuclease I - chemistry
/ Deoxyribonuclease I - drug effects
/ Deoxyribonuclease I - radiation effects
/ DNA
/ EDTA
/ Electrophoresis, Polyacrylamide Gel
/ Endangered & extinct species
/ Enzymes
/ Medicine
/ Medicine and Health Sciences
/ Methods
/ Nuclease
/ Peptide Fragments - analysis
/ Proteins
/ Proteolysis - radiation effects
/ Research and analysis methods
/ Ribonuclease, Pancreatic - chemistry
/ Ribonuclease, Pancreatic - drug effects
/ Ribonuclease, Pancreatic - radiation effects
/ Ribonucleases - drug effects
/ Ribonucleases - radiation effects
/ RNA
