Laboratory Exercise to Measure Plasmid Copy Number by qPCR

ABSTRACT 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,...

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Autores principales:	Benjamin David, Jinbei Li, Faisal Masood, Caroline Blassick, Paul Jensen, Karin Jensen
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2021
Materias:	qPCR laboratory instruction Special aspects of education LC8-6691 Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/0df22c06cf304ff1b00369fad7640d6c
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spelling	oai:doaj.org-article:0df22c06cf304ff1b00369fad7640d6c2021-11-15T15:04:52ZLaboratory Exercise to Measure Plasmid Copy Number by qPCR10.1128/jmbe.00125-211935-78851935-7877https://doaj.org/article/0df22c06cf304ff1b00369fad7640d6c2021-09-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/jmbe.00125-21https://doaj.org/toc/1935-7877https://doaj.org/toc/1935-7885ABSTRACT 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 (CT) 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).Benjamin DavidJinbei LiFaisal MasoodCaroline BlassickPaul JensenKarin JensenAmerican Society for MicrobiologyarticleqPCRlaboratory instructionSpecial aspects of educationLC8-6691Biology (General)QH301-705.5ENJournal of Microbiology & Biology Education, Vol 22, Iss 2 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	qPCR laboratory instruction Special aspects of education LC8-6691 Biology (General) QH301-705.5
spellingShingle	qPCR laboratory instruction Special aspects of education LC8-6691 Biology (General) QH301-705.5 Benjamin David Jinbei Li Faisal Masood Caroline Blassick Paul Jensen Karin Jensen Laboratory Exercise to Measure Plasmid Copy Number by qPCR
description	ABSTRACT 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 (CT) 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	article
author	Benjamin David Jinbei Li Faisal Masood Caroline Blassick Paul Jensen Karin Jensen
author_facet	Benjamin David Jinbei Li Faisal Masood Caroline Blassick Paul Jensen Karin Jensen
author_sort	Benjamin David
title	Laboratory Exercise to Measure Plasmid Copy Number by qPCR
title_short	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_sort	laboratory exercise to measure plasmid copy number by qpcr
publisher	American Society for Microbiology
publishDate	2021
url	https://doaj.org/article/0df22c06cf304ff1b00369fad7640d6c
work_keys_str_mv	AT benjamindavid laboratoryexercisetomeasureplasmidcopynumberbyqpcr AT jinbeili laboratoryexercisetomeasureplasmidcopynumberbyqpcr AT faisalmasood laboratoryexercisetomeasureplasmidcopynumberbyqpcr AT carolineblassick laboratoryexercisetomeasureplasmidcopynumberbyqpcr AT pauljensen laboratoryexercisetomeasureplasmidcopynumberbyqpcr AT karinjensen laboratoryexercisetomeasureplasmidcopynumberbyqpcr
_version_	1718428172923437056

Laboratory Exercise to Measure Plasmid Copy Number by qPCR

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