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Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies
[Image: see text] Microfluidic platforms offer a drastic increase in throughput while minimizing sample usage and hands-on time, which make them important tools for large-scale biological studies. A range of such systems have been developed for enzyme activity studies, although their complexity larg...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Chemical Society
2021
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7841792/ https://www.ncbi.nlm.nih.gov/pubmed/33521438 http://dx.doi.org/10.1021/acsomega.0c04918 |
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author | Rembeza, Elzbieta Engqvist, Martin K. M. |
author_facet | Rembeza, Elzbieta Engqvist, Martin K. M. |
author_sort | Rembeza, Elzbieta |
collection | PubMed |
description | [Image: see text] Microfluidic platforms offer a drastic increase in throughput while minimizing sample usage and hands-on time, which make them important tools for large-scale biological studies. A range of such systems have been developed for enzyme activity studies, although their complexity largely hinders their application to the wider scientific community. Here, we present adaptation of an easy-to-use commercial microfluidic qPCR system for performing enzyme kinetic studies. We demonstrate the functionality of the Fluidigm Biomark HD system (the Fluidigm system) by determining the kinetic properties of three oxidases in a resorufin-based fluorescence assay. The results obtained in the microfluidic system proved reproducible and comparable to the ones obtained in a standard microplate-based assay. With a wide range of easy-to-use, off-the-shelf components, the microfluidic system presents itself as a simple and customizable platform for high-throughput enzyme activity studies. |
format | Online Article Text |
id | pubmed-7841792 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Chemical Society |
record_format | MEDLINE/PubMed |
spelling | pubmed-78417922021-01-29 Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies Rembeza, Elzbieta Engqvist, Martin K. M. ACS Omega [Image: see text] Microfluidic platforms offer a drastic increase in throughput while minimizing sample usage and hands-on time, which make them important tools for large-scale biological studies. A range of such systems have been developed for enzyme activity studies, although their complexity largely hinders their application to the wider scientific community. Here, we present adaptation of an easy-to-use commercial microfluidic qPCR system for performing enzyme kinetic studies. We demonstrate the functionality of the Fluidigm Biomark HD system (the Fluidigm system) by determining the kinetic properties of three oxidases in a resorufin-based fluorescence assay. The results obtained in the microfluidic system proved reproducible and comparable to the ones obtained in a standard microplate-based assay. With a wide range of easy-to-use, off-the-shelf components, the microfluidic system presents itself as a simple and customizable platform for high-throughput enzyme activity studies. American Chemical Society 2021-01-13 /pmc/articles/PMC7841792/ /pubmed/33521438 http://dx.doi.org/10.1021/acsomega.0c04918 Text en © 2021 The Authors. Published by American Chemical Society This is an open access article published under a Creative Commons Attribution (CC-BY) License (http://pubs.acs.org/page/policy/authorchoice_ccby_termsofuse.html) , which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited. |
spellingShingle | Rembeza, Elzbieta Engqvist, Martin K. M. Adaptation of a Microfluidic qPCR System for Enzyme Kinetic Studies |
title | Adaptation of a Microfluidic qPCR System for Enzyme
Kinetic Studies |
title_full | Adaptation of a Microfluidic qPCR System for Enzyme
Kinetic Studies |
title_fullStr | Adaptation of a Microfluidic qPCR System for Enzyme
Kinetic Studies |
title_full_unstemmed | Adaptation of a Microfluidic qPCR System for Enzyme
Kinetic Studies |
title_short | Adaptation of a Microfluidic qPCR System for Enzyme
Kinetic Studies |
title_sort | adaptation of a microfluidic qpcr system for enzyme
kinetic studies |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7841792/ https://www.ncbi.nlm.nih.gov/pubmed/33521438 http://dx.doi.org/10.1021/acsomega.0c04918 |
work_keys_str_mv | AT rembezaelzbieta adaptationofamicrofluidicqpcrsystemforenzymekineticstudies AT engqvistmartinkm adaptationofamicrofluidicqpcrsystemforenzymekineticstudies |