The protocol presented here allows the isolation, purification, nucleic acid extraction, and amplification of DNA/RNA from viruses present in human sera samples. The method allows the random amplification of the viral genomes present by using a Sequence-Independent, Single-Primer Amplification (SISPA) approach enabling the study of both DNA/RNA viruses. An amplification step is needed, as the concentration of viral DNA/RNA in serum samples is low for direct library preparation. The application of the described protocol guarantees enough randomly amplified double-strand DNA for further library preparation using Nextera XT kit from Illumina.
CITATION STYLE
Fernandez-Cassi, X., Rusiñol, M., & Martínez-Puchol, S. (2018). Viral concentration and amplification from human serum samples prior to application of next-generation sequencing analysis. In Methods in Molecular Biology (Vol. 1838, pp. 173–188). Humana Press Inc. https://doi.org/10.1007/978-1-4939-8682-8_13
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