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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...

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Detalles Bibliográficos
Autores principales: Kachkin, Daniel V., Khorolskaya, Julia I., Ivanova, Julia S., Rubel, Aleksandr A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
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.
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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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