Figure 2.

Verification of Cam^r/Zeo^s/Suc^r colonies for the presence of U_L34 recombinant HSV-BAC clones by PCR. (A and B) Photographs of electrophoretically separated PCR products amplified directly from positive colonies. (C) Restriction digests of the PCR products amplified from the potential positive colonies for U_L34Δ(245–275). The PCR product was clarified by Gene-clean (Biol01) and digested with PacI, a restriction site expected if the integration of pRB5715 had taken place. RH18 1–4, RH20 1–4, and RH17 1–4 represent individual colonies selected for analysis. Marker, 1-kb ladder of molecular weight marker. The arrows identify the PCR-positive clones, which gave amplified DNA band of expected size or were cleaved by the appropriate restriction endonuclease.