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pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis

DNA size markers (also known as ‘molecular weight markers’ or ‘DNA ladders’) are an essential tool when using gel electrophoresis to identify and purify nucleic acids. However, the cost of these DNA ladders is not insignificant and, over time, impinges on the funds available for research and trainin...

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Detalles Bibliográficos
Autores principales: Allen, Angel G, Barthelson, Karissa, Lardelli, Michael
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10400483/
https://www.ncbi.nlm.nih.gov/pubmed/37547169
http://dx.doi.org/10.1093/biomethods/bpad015
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author Allen, Angel G
Barthelson, Karissa
Lardelli, Michael
author_facet Allen, Angel G
Barthelson, Karissa
Lardelli, Michael
author_sort Allen, Angel G
collection PubMed
description DNA size markers (also known as ‘molecular weight markers’ or ‘DNA ladders’) are an essential tool when using gel electrophoresis to identify and purify nucleic acids. However, the cost of these DNA ladders is not insignificant and, over time, impinges on the funds available for research and training in molecular biology. Here, we describe a method for the generation of ‘pHAPE’, a plasmid from which a variety of DNA ladders can be generated via simple restriction enzyme digestions. The pHAPE plasmid can be generated by mutagenesis of the commonly used pBluescript II SK+ phagemid followed by insertion of a 7141 bp sequence (comprised of three smaller, synthetic fragments). Our use of pHAPE allows us some small relief from the ever-rising costs of performing molecular biology experiments (‘Don’t worry, pHAPE’).
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spelling pubmed-104004832023-08-05 pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis Allen, Angel G Barthelson, Karissa Lardelli, Michael Biol Methods Protoc Methods Article DNA size markers (also known as ‘molecular weight markers’ or ‘DNA ladders’) are an essential tool when using gel electrophoresis to identify and purify nucleic acids. However, the cost of these DNA ladders is not insignificant and, over time, impinges on the funds available for research and training in molecular biology. Here, we describe a method for the generation of ‘pHAPE’, a plasmid from which a variety of DNA ladders can be generated via simple restriction enzyme digestions. The pHAPE plasmid can be generated by mutagenesis of the commonly used pBluescript II SK+ phagemid followed by insertion of a 7141 bp sequence (comprised of three smaller, synthetic fragments). Our use of pHAPE allows us some small relief from the ever-rising costs of performing molecular biology experiments (‘Don’t worry, pHAPE’). Oxford University Press 2023-07-31 /pmc/articles/PMC10400483/ /pubmed/37547169 http://dx.doi.org/10.1093/biomethods/bpad015 Text en © The Author(s) 2023. Published by Oxford University Press. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com
spellingShingle Methods Article
Allen, Angel G
Barthelson, Karissa
Lardelli, Michael
pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis
title pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis
title_full pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis
title_fullStr pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis
title_full_unstemmed pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis
title_short pHAPE: a plasmid for production of DNA size marker ladders for gel electrophoresis
title_sort phape: a plasmid for production of dna size marker ladders for gel electrophoresis
topic Methods Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10400483/
https://www.ncbi.nlm.nih.gov/pubmed/37547169
http://dx.doi.org/10.1093/biomethods/bpad015
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