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An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures
In this article, we present several protocols that describe the steps from cloning and obtaining a large amount of pure plasmid DNA to generation of lentiviruses based on these constructs. The protocols have been worked out on human cell culture HEK293T but can be adapted for other cell cultures. Th...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7712542/ https://www.ncbi.nlm.nih.gov/pubmed/33066602 http://dx.doi.org/10.3390/mps3040069 |
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author | Kachkin, Daniel V. Khorolskaya, Julia I. Ivanova, Julia S. Rubel, Aleksandr A. |
author_facet | Kachkin, Daniel V. Khorolskaya, Julia I. Ivanova, Julia S. Rubel, Aleksandr A. |
author_sort | Kachkin, Daniel V. |
collection | PubMed |
description | In this article, we present several protocols that describe the steps from cloning and obtaining a large amount of pure plasmid DNA to generation of lentiviruses based on these constructs. The protocols have been worked out on human cell culture HEK293T but can be adapted for other cell cultures. This protocol was designed to be simple to execute and cheap since it requires only materials and consumables widely available in molecular laboratories, such as salts, alcohols, etc., and no complicated laboratory equipment. These protocols are highly effective and can be performed in any standard molecular biology laboratory. |
format | Online Article Text |
id | pubmed-7712542 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-77125422020-12-04 An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures Kachkin, Daniel V. Khorolskaya, Julia I. Ivanova, Julia S. Rubel, Aleksandr A. Methods Protoc Protocol In this article, we present several protocols that describe the steps from cloning and obtaining a large amount of pure plasmid DNA to generation of lentiviruses based on these constructs. The protocols have been worked out on human cell culture HEK293T but can be adapted for other cell cultures. This protocol was designed to be simple to execute and cheap since it requires only materials and consumables widely available in molecular laboratories, such as salts, alcohols, etc., and no complicated laboratory equipment. These protocols are highly effective and can be performed in any standard molecular biology laboratory. MDPI 2020-10-14 /pmc/articles/PMC7712542/ /pubmed/33066602 http://dx.doi.org/10.3390/mps3040069 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Kachkin, Daniel V. Khorolskaya, Julia I. Ivanova, Julia S. Rubel, Aleksandr A. An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures |
title | An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures |
title_full | An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures |
title_fullStr | An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures |
title_full_unstemmed | An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures |
title_short | An Efficient Method for Isolation of Plasmid DNA for Transfection of Mammalian Cell Cultures |
title_sort | efficient method for isolation of plasmid dna for transfection of mammalian cell cultures |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7712542/ https://www.ncbi.nlm.nih.gov/pubmed/33066602 http://dx.doi.org/10.3390/mps3040069 |
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