Fig. (2).

Multiple displacement amplification (MDA) – single HIV-1 genome sequencing (SGS) workflow. Genomic PBMC DNA from an HIV-infected individual is diluted and dispensed into 96-well plates to maximize the number of wells containing exactly one provirus. MDA is then used to amplify DNA in each of these wells on the order of 1000-fold. Reaction products from wells originally containing a single HIV-1 provirus may thus be partitioned for downstream assays, including PCR-amplification and sequencing of segments of the virus genome (e.g., the RT gene), amplification and sequencing of near full length (NFL) provirus, and integration site determination. Further and more complete characterization of the latent HIV-1 reservoir will depend upon continued development of techniques such as these that permit examination of multiple facets of individual HIV-infected cells.