Affiliation:
1. CIC bioGUNE, Bizkaia Science and Technology Park, Building 800, 48160 Derio, Spain
2. Department of Biochemistry, Universidad Autónoma de Madrid and Instituto de Investigaciones Biomédicas “Alberto Sols”, CSIC-UAM, 28029 Madrid, Spain
Abstract
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.
Subject
Inorganic Chemistry,Organic Chemistry,Physical and Theoretical Chemistry,Computer Science Applications,Spectroscopy,Molecular Biology,General Medicine,Catalysis
Reference183 articles.
1. DNA Polymerases and Biotechnological Applications;Aschenbrenner;Curr. Opin. Biotechnol.,2017
2. Benchmarking of Single-Virus Genomics: A New Tool for Uncovering the Virosphere;Bhattacharjee;Environ. Microbiol.,2021
3. Khan, T., Becker, T.M., Po, J.W., Chua, W., and Ma, Y. (2022). Single-Circulating Tumor Cell Whole Genome Amplification to Unravel Cancer Heterogeneity and Actionable Biomarkers. Int. J. Mol. Sci., 23.
4. Construction of Small-Insert and Large-Insert Metagenomic Libraries;Simon;Methods Mol. Biol. Clifton NJ,2023
5. Principles of Whole-Genome Amplification;Czyz;Methods Mol. Biol.,2015
Cited by
8 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献