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Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits

Silica columns from PCR purification and gel extraction kits are widely used in laboratories worldwide to assist in gene cloning. However, the use of these columns can generate plastic waste that has an environmental impact due to their one-off design and massive consumption. Thus, it is important t...

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Autores principales: Zhou, Ying, Zhang, Yang, He, Wei, Wang, Juan, Peng, Feixia, Huang, Liyun, Zhao, Shasha, Deng, Wensheng
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6110862/
https://www.ncbi.nlm.nih.gov/pubmed/30150610
http://dx.doi.org/10.1038/s41598-018-30316-w
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author Zhou, Ying
Zhang, Yang
He, Wei
Wang, Juan
Peng, Feixia
Huang, Liyun
Zhao, Shasha
Deng, Wensheng
author_facet Zhou, Ying
Zhang, Yang
He, Wei
Wang, Juan
Peng, Feixia
Huang, Liyun
Zhao, Shasha
Deng, Wensheng
author_sort Zhou, Ying
collection PubMed
description Silica columns from PCR purification and gel extraction kits are widely used in laboratories worldwide to assist in gene cloning. However, the use of these columns can generate plastic waste that has an environmental impact due to their one-off design and massive consumption. Thus, it is important to develop a novel method that can reduce the utilization of silica columns but not affect research efficiency. In this study, various chemical and nonchemical reagents were used to eliminate residual DNA within used columns from PCR purification and gel extraction kits. We show that phosphoric acid is the most effective reagent among those tested to remove DNA contamination from used columns. Columns regenerated using 1 M phosphoric acid have a DNA purification capability that is comparable to that of fresh columns. We demonstrate that silica columns can be regenerated and reused a minimum of five times. The lab-made buffers are compatible with the regenerated columns for DNA purification, and DNA that is prepared with the regenerated columns can be used for gene cloning without affecting the gene cloning efficiency. Thus, the use of this novel method greatly reduces the production of laboratory waste and benefits numerous laboratories worldwide.
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spelling pubmed-61108622018-08-30 Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits Zhou, Ying Zhang, Yang He, Wei Wang, Juan Peng, Feixia Huang, Liyun Zhao, Shasha Deng, Wensheng Sci Rep Article Silica columns from PCR purification and gel extraction kits are widely used in laboratories worldwide to assist in gene cloning. However, the use of these columns can generate plastic waste that has an environmental impact due to their one-off design and massive consumption. Thus, it is important to develop a novel method that can reduce the utilization of silica columns but not affect research efficiency. In this study, various chemical and nonchemical reagents were used to eliminate residual DNA within used columns from PCR purification and gel extraction kits. We show that phosphoric acid is the most effective reagent among those tested to remove DNA contamination from used columns. Columns regenerated using 1 M phosphoric acid have a DNA purification capability that is comparable to that of fresh columns. We demonstrate that silica columns can be regenerated and reused a minimum of five times. The lab-made buffers are compatible with the regenerated columns for DNA purification, and DNA that is prepared with the regenerated columns can be used for gene cloning without affecting the gene cloning efficiency. Thus, the use of this novel method greatly reduces the production of laboratory waste and benefits numerous laboratories worldwide. Nature Publishing Group UK 2018-08-27 /pmc/articles/PMC6110862/ /pubmed/30150610 http://dx.doi.org/10.1038/s41598-018-30316-w Text en © The Author(s) 2018 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Zhou, Ying
Zhang, Yang
He, Wei
Wang, Juan
Peng, Feixia
Huang, Liyun
Zhao, Shasha
Deng, Wensheng
Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits
title Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits
title_full Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits
title_fullStr Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits
title_full_unstemmed Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits
title_short Rapid Regeneration and Reuse of Silica Columns from PCR Purification and Gel Extraction Kits
title_sort rapid regeneration and reuse of silica columns from pcr purification and gel extraction kits
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6110862/
https://www.ncbi.nlm.nih.gov/pubmed/30150610
http://dx.doi.org/10.1038/s41598-018-30316-w
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