Figure 1

Methods to remove RTIs from virus preparations. Schematic of the two methods used to remove RTIs from virus preparations. The RTIs were removed so that they did not inhibit downstream assays to assess viral function when premature reverse transcription was blocked. In both methods aspiration was used to remove the supernatant after centrifugation (see the Methods section for details). The method on the left was used to i) maintain competent Env proteins on the surface of virions and ii) limit mechanical stress on virions for subsequent infection analyses. The method on the right uses DNase I treatment to remove extra-virion plasmid DNA contamination with subsequent subtilisin digestion to ensure that the DNase I is completely removed prior to lysis of the virions, and qPCR analysis of intravirion DNA and endogenous reverse transcription assays [33].