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Laboratory Exercise to Measure Plasmid Copy Number by qPCR
Quantitative PCR (qPCR) has numerous applications in biology. In an educational setting, qPCR provides students an opportunity to better understand the PCR mechanism by providing both quantitative information about the reactions and also data to troubleshoot PCRs (e.g., melt curves). Here, we presen...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
American Society for Microbiology
2021
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8442022/ https://www.ncbi.nlm.nih.gov/pubmed/34594455 http://dx.doi.org/10.1128/jmbe.00125-21 |
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author | David, Benjamin Li, Jinbei Masood, Faisal Blassick, Caroline Jensen, Paul Jensen, Karin |
author_facet | David, Benjamin Li, Jinbei Masood, Faisal Blassick, Caroline Jensen, Paul Jensen, Karin |
author_sort | David, Benjamin |
collection | PubMed |
description | Quantitative PCR (qPCR) has numerous applications in biology. In an educational setting, qPCR provides students an opportunity to better understand the PCR mechanism by providing both quantitative information about the reactions and also data to troubleshoot PCRs (e.g., melt curves). Here, we present a relatively short (2-h) laboratory activity to demonstrate qPCR to quantify plasmid copy number (CN) by measuring the cycle threshold (C(T)) values for a genomic gene and a plasmid gene using transformed cells as a template. The activity can be combined with additional laboratory exercises, including bacterial transformation, to create the template to be used in the qPCRs. This lab activity is ideal for undergraduate laboratory courses that include recombinant DNA technology. (This work was presented at the 2020 Biomedical Engineering Society annual meeting). |
format | Online Article Text |
id | pubmed-8442022 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | American Society for Microbiology |
record_format | MEDLINE/PubMed |
spelling | pubmed-84420222021-09-29 Laboratory Exercise to Measure Plasmid Copy Number by qPCR David, Benjamin Li, Jinbei Masood, Faisal Blassick, Caroline Jensen, Paul Jensen, Karin J Microbiol Biol Educ Tips and Tools Quantitative PCR (qPCR) has numerous applications in biology. In an educational setting, qPCR provides students an opportunity to better understand the PCR mechanism by providing both quantitative information about the reactions and also data to troubleshoot PCRs (e.g., melt curves). Here, we present a relatively short (2-h) laboratory activity to demonstrate qPCR to quantify plasmid copy number (CN) by measuring the cycle threshold (C(T)) values for a genomic gene and a plasmid gene using transformed cells as a template. The activity can be combined with additional laboratory exercises, including bacterial transformation, to create the template to be used in the qPCRs. This lab activity is ideal for undergraduate laboratory courses that include recombinant DNA technology. (This work was presented at the 2020 Biomedical Engineering Society annual meeting). American Society for Microbiology 2021-07-30 /pmc/articles/PMC8442022/ /pubmed/34594455 http://dx.doi.org/10.1128/jmbe.00125-21 Text en Copyright © 2021 David et al. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Tips and Tools David, Benjamin Li, Jinbei Masood, Faisal Blassick, Caroline Jensen, Paul Jensen, Karin Laboratory Exercise to Measure Plasmid Copy Number by qPCR |
title | Laboratory Exercise to Measure Plasmid Copy Number by qPCR |
title_full | Laboratory Exercise to Measure Plasmid Copy Number by qPCR |
title_fullStr | Laboratory Exercise to Measure Plasmid Copy Number by qPCR |
title_full_unstemmed | Laboratory Exercise to Measure Plasmid Copy Number by qPCR |
title_short | Laboratory Exercise to Measure Plasmid Copy Number by qPCR |
title_sort | laboratory exercise to measure plasmid copy number by qpcr |
topic | Tips and Tools |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8442022/ https://www.ncbi.nlm.nih.gov/pubmed/34594455 http://dx.doi.org/10.1128/jmbe.00125-21 |
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