Laboratory Exercise To Measure Restriction Enzyme Kinetics

Enzymes are ubiquitous in the fields of biology and microbiology, catalyzing critical reactions and enabling a broad range of biotechnological applications. Despite the important role that enzyme catalysis plays in biological processes, undergraduate students often struggle to understand enzyme kine...

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Autores principales:	Caroline Blassick, Benjamin David, Audra Storm, Paul Jensen, Karin Jensen
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2019
Materias:	Special aspects of education LC8-6691 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/c77404ba5d594d85946c4bc823f2e59a
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spelling	oai:doaj.org-article:c77404ba5d594d85946c4bc823f2e59a2021-11-15T15:04:12ZLaboratory Exercise To Measure Restriction Enzyme Kinetics10.1128/jmbe.v20i3.17031935-78851935-7877https://doaj.org/article/c77404ba5d594d85946c4bc823f2e59a2019-01-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/jmbe.v20i3.1703https://doaj.org/toc/1935-7877https://doaj.org/toc/1935-7885Enzymes are ubiquitous in the fields of biology and microbiology, catalyzing critical reactions and enabling a broad range of biotechnological applications. Despite the important role that enzyme catalysis plays in biological processes, undergraduate students often struggle to understand enzyme kinetics in the classroom. In an attempt to improve students’ understanding of the topic, we present a relatively short and inexpensive laboratory activity designed to give students hands-on experience with generating and manipulating enzyme kinetic data. Students perform restriction digests of DNA at various time points, visualize the reaction products on an agarose gel, and quantify their data in order to construct Lineweaver-Burk plots which compare the effects of a restriction enzyme and its engineered version. The activity may be completed in a single two-hour lab session and, unlike other enzyme assays designed for laboratory courses, does not require a microplate reader to complete. The activity allows students to see connections between a visual data set and quantitative kinetic data, in order to solidify their understanding of enzyme kinetics. Students also learn the skills of gel electrophoresis and image quantification using ImageJ software. This lab activity is ideal for undergraduate laboratory courses which address enzyme kinetics and DNA technology.Caroline BlassickBenjamin DavidAudra StormPaul JensenKarin JensenAmerican Society for MicrobiologyarticleSpecial aspects of educationLC8-6691Biology (General)QH301-705.5ENJournal of Microbiology & Biology Education, Vol 20, Iss 3 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Special aspects of education LC8-6691 Biology (General) QH301-705.5
spellingShingle	Special aspects of education LC8-6691 Biology (General) QH301-705.5 Caroline Blassick Benjamin David Audra Storm Paul Jensen Karin Jensen Laboratory Exercise To Measure Restriction Enzyme Kinetics
description	Enzymes are ubiquitous in the fields of biology and microbiology, catalyzing critical reactions and enabling a broad range of biotechnological applications. Despite the important role that enzyme catalysis plays in biological processes, undergraduate students often struggle to understand enzyme kinetics in the classroom. In an attempt to improve students’ understanding of the topic, we present a relatively short and inexpensive laboratory activity designed to give students hands-on experience with generating and manipulating enzyme kinetic data. Students perform restriction digests of DNA at various time points, visualize the reaction products on an agarose gel, and quantify their data in order to construct Lineweaver-Burk plots which compare the effects of a restriction enzyme and its engineered version. The activity may be completed in a single two-hour lab session and, unlike other enzyme assays designed for laboratory courses, does not require a microplate reader to complete. The activity allows students to see connections between a visual data set and quantitative kinetic data, in order to solidify their understanding of enzyme kinetics. Students also learn the skills of gel electrophoresis and image quantification using ImageJ software. This lab activity is ideal for undergraduate laboratory courses which address enzyme kinetics and DNA technology.
format	article
author	Caroline Blassick Benjamin David Audra Storm Paul Jensen Karin Jensen
author_facet	Caroline Blassick Benjamin David Audra Storm Paul Jensen Karin Jensen
author_sort	Caroline Blassick
title	Laboratory Exercise To Measure Restriction Enzyme Kinetics
title_short	Laboratory Exercise To Measure Restriction Enzyme Kinetics
title_full	Laboratory Exercise To Measure Restriction Enzyme Kinetics
title_fullStr	Laboratory Exercise To Measure Restriction Enzyme Kinetics
title_full_unstemmed	Laboratory Exercise To Measure Restriction Enzyme Kinetics
title_sort	laboratory exercise to measure restriction enzyme kinetics
publisher	American Society for Microbiology
publishDate	2019
url	https://doaj.org/article/c77404ba5d594d85946c4bc823f2e59a
work_keys_str_mv	AT carolineblassick laboratoryexercisetomeasurerestrictionenzymekinetics AT benjamindavid laboratoryexercisetomeasurerestrictionenzymekinetics AT audrastorm laboratoryexercisetomeasurerestrictionenzymekinetics AT pauljensen laboratoryexercisetomeasurerestrictionenzymekinetics AT karinjensen laboratoryexercisetomeasurerestrictionenzymekinetics
_version_	1718428275971194880

Laboratory Exercise To Measure Restriction Enzyme Kinetics

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