FIG. 2.

HIV-1 sequence-specific UDG cloning. In this schematic representation of HIV-1 sequence-specific UDG cloning, pJM20ΔGPRT is inverse PCR amplified and the vector PCR product is directly mixed and annealed with the Gag p7/p1 and Gag p1/p6 cleavage site, PR, and RT (GPRT) insert PCR amplicon obtained from HIV-1 RNA. After the UDG digestion, the 5′ ends of both PCR products become long sticky ends, up to the last dUMP incorporated in the primer, and stably anneal. The mix is then electroporated into E. coli, where gaps are repaired. The white arrowhead indicates the restriction site used to linearize pJM20ΔGPRT before inverse vector PCR.