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Cloning of the IS. Primers C-EBNA1-1 and C-EBNA1-2 were designed with 23 randomized bases at the 5′ end. Two separate PCRs with primers C-EBNA1-2 and QP1 and primers C-EBNA1-1 and QP2 were performed. Purified PCR products were mixed and reamplified with primers QP1 and QP2. A BamHI K rightward frame 1 (BKRF1) PCR product with 23 internally randomized bases compared to the wild-type gene was obtained and cloned into pCR-Script SK(+), giving pQPCR-8.
