Publication: Comparison of phi29-based whole genome amplification and whole transcriptome amplification in dengue virus
Issued Date
2014-01-01
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ISSN
18790984
01660934
01660934
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2-s2.0-84887260829
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Mahidol University
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SCOPUS
Bibliographic Citation
Journal of Virological Methods. Vol.195, (2014), 141-147
Suggested Citation
Patcharawan Sujayanont, Kwanrutai Chininmanu, Boonrat Tassaneetrithep, Nattaya Tangthawornchaikul, Prida Malasit, Prapat Suriyaphol Comparison of phi29-based whole genome amplification and whole transcriptome amplification in dengue virus. Journal of Virological Methods. Vol.195, (2014), 141-147. doi:10.1016/j.jviromet.2013.10.005 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/34019
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Title
Comparison of phi29-based whole genome amplification and whole transcriptome amplification in dengue virus
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Abstract
Dengue virus is responsible for 50-100 million new infections annually worldwide. The virus uses error-prone RNA polymerase during genome replication in a host, resulting in the formation of closely related viruses known as quasispecies. The availability of next-generation sequencing technology provides opportunities to analyze viral quasispecies. Before analysis, it is crucial to increase the amount of DNA because of the limited amounts of viral genomic material that can be isolated from a patient. However, using specific primers may overlook the occurrence of possible variations at primer binding sites. To address this problem, the performance of two sequence-independent amplification methods was compared for whole genome amplification (WGA): phi29 DNA polymerase-based WGA and whole transcriptome amplification (WTA). Both methods have the ability to provide complete coverage of the dengue genome from template amounts as low as 1. ng. However, WTA showed greater efficiency in terms of yield (WTA: ~10. μg; phi29-based WGA: ~500. ng) and lower amplification bias. In conclusion, the WTA amplification kit was shown to perform substantially better than phi29 DNA polymerase-based WGA in terms of both final concentration and amplification bias in amplifying small genomes, such as that of the dengue virus. © 2013 The Authors.