An experimental strategy for preparing circular ssDNA virus genomes for next-generation sequencing

Feb 2022
Authors:
Catherine D. Aimone, Steen Hoyer, Anna E. Dye, David O. Deppong, Siobain Duffy, Ignazio Carbone, Linda Hanley-Bowdoin

Info:
The authors provide a protocol for analyzing single stranded DNA (ssDNA) from begomoviruses, which cause significant damage to many crops (e.g. cassava and tomato), and is transmitted by whiteflies. The ssDNA is enriched from both plants and whiteflies and combined rolling circle amplification (RCA) and DNA size selection to prepare samples for sequencing and analysis of the viral genomes. The authors suggest that this method can be used to examine viral DNA as it moves from host to vector and be used for viral DNA population studies.
BluePippin was used for DNA size selection.

Author Affiliations:
Department of Plant and Microbial Biology, North Carolina State University, Raleigh, NC,
Department of Ecology, Evolution, and Natural Resources, Rutgers University, New Brunswick, NJ
Center for Integrated Fungal Research, Department of Entomology and Plant Pathology, North Carolina State University, Raleigh, NC

Journal of Virological Methods
DOI: 10.1016/j.jviromet.2021.114405

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