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

Jun 1, 2022

Cassava and tomato are important crops, whose processing market reached a volume of 298.8 and 39.2 million metric tons in 2020, respectively (Council., 2021; imarc, 2021). Yet, both cassava and tomato yields are severely impacted by begomovirus diseases. Begomoviruses are single-stranded DNA (ssDNA) viruses that are transmitted by whiteflies (Bemisia tabaci), and consist of either a single DNA component (monopartite) or two DNA components (bipartite), designated as DNA-A and DNA-B. The total DNA from cassava, tomato and whiteflies (250 ng) was selected for 1-6 Kb DNA size fragments by electrophoresis on a 0.75 % agarose gel at 25V DC for 3–8 h using the Blue Pippin Prep system (Model # BDQ3010, Sage Science, Beverly MA). BluePippin system was used to select for DNA < 6 Kb from total DNA samples isolated from symptomatic Kibandameno leaves from two plants.

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Preparative electrophoresis systems for targeted size selection of biomolecules.

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