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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
Descripción
Sumario: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’).