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DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions
In the same way that specialized DNA polymerases (DNAPs) replicate cellular and viral genomes, only a handful of dedicated proteins from various natural origins as well as engineered versions are appropriate for competent exponential amplification of whole genomes and metagenomes (WGA). Different ap...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10253169/ https://www.ncbi.nlm.nih.gov/pubmed/37298280 http://dx.doi.org/10.3390/ijms24119331 |
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author | Ordóñez, Carlos D. Redrejo-Rodríguez, Modesto |
author_facet | Ordóñez, Carlos D. Redrejo-Rodríguez, Modesto |
author_sort | Ordóñez, Carlos D. |
collection | PubMed |
description | In the same way that specialized DNA polymerases (DNAPs) replicate cellular and viral genomes, only a handful of dedicated proteins from various natural origins as well as engineered versions are appropriate for competent exponential amplification of whole genomes and metagenomes (WGA). Different applications have led to the development of diverse protocols, based on various DNAPs. Isothermal WGA is currently widely used due to the high performance of Φ29 DNA polymerase, but PCR-based methods are also available and can provide competent amplification of certain samples. Replication fidelity and processivity must be considered when selecting a suitable enzyme for WGA. However, other properties, such as thermostability, capacity to couple replication, and double helix unwinding, or the ability to maintain DNA replication opposite to damaged bases, are also very relevant for some applications. In this review, we provide an overview of the different properties of DNAPs widely used in WGA and discuss their limitations and future research directions. |
format | Online Article Text |
id | pubmed-10253169 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-102531692023-06-10 DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions Ordóñez, Carlos D. Redrejo-Rodríguez, Modesto Int J Mol Sci Review In the same way that specialized DNA polymerases (DNAPs) replicate cellular and viral genomes, only a handful of dedicated proteins from various natural origins as well as engineered versions are appropriate for competent exponential amplification of whole genomes and metagenomes (WGA). Different applications have led to the development of diverse protocols, based on various DNAPs. Isothermal WGA is currently widely used due to the high performance of Φ29 DNA polymerase, but PCR-based methods are also available and can provide competent amplification of certain samples. Replication fidelity and processivity must be considered when selecting a suitable enzyme for WGA. However, other properties, such as thermostability, capacity to couple replication, and double helix unwinding, or the ability to maintain DNA replication opposite to damaged bases, are also very relevant for some applications. In this review, we provide an overview of the different properties of DNAPs widely used in WGA and discuss their limitations and future research directions. MDPI 2023-05-26 /pmc/articles/PMC10253169/ /pubmed/37298280 http://dx.doi.org/10.3390/ijms24119331 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/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 (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Review Ordóñez, Carlos D. Redrejo-Rodríguez, Modesto DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions |
title | DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions |
title_full | DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions |
title_fullStr | DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions |
title_full_unstemmed | DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions |
title_short | DNA Polymerases for Whole Genome Amplification: Considerations and Future Directions |
title_sort | dna polymerases for whole genome amplification: considerations and future directions |
topic | Review |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10253169/ https://www.ncbi.nlm.nih.gov/pubmed/37298280 http://dx.doi.org/10.3390/ijms24119331 |
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