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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...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2023
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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’). |
format | Online Article Text |
id | pubmed-10400483 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
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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