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. 2021 May 10;93(20):7422–7429. doi: 10.1021/acs.analchem.0c04749

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© 2021 The Authors. Published by American Chemical Society

Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).

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Compound enrichment and sequencing of HIV proviruses in infected individuals. (a) The first round of NAC uses a TaqMan assay targeting the pol gene (FAM). The second round of NAC uses a degenerate TaqMan assay targeting the long terminal repeat (LTR, FAM) and tat gene (Cy5). (b) Implementation of in-droplet MDA before the second round generates sufficient material for single droplet sequencing. The MDA droplet is merged with a droplet containing PCR reagents for TaqMan detection in the second round. (c) HIV genome and integration site coverage maps for nonenriched sample (top panel) and three sorted individual drops. Aggregating all data (bottom panel) assembles the full-length HIV genome including the human integration site. The peaks at LTR and tat are the TaqMan PCR amplicons.