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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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Autores principales: Ordóñez, Carlos D., Redrejo-Rodríguez, Modesto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
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.
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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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