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. Author manuscript; available in PMC: 2017 Jun 2.
Published in final edited form as: Virus Res. 2016 Mar 26;217:107–114. doi: 10.1016/j.virusres.2016.02.011

Figure 2.

Figure 2

Recombination cassette assembly. (A.) PCR amplification of the LA, RA and CMV-GNR products (lanes 2, 3 and 4, respectively). (B.) Assembly by overlapping PCR. Individual PCR products for the LA, RA and CMV-GNR are combined and assembled using a standard PCR protocol. Lane 1, molecular weight marker, and lanes 2 – 4 show the assembly of recombination cassettes to delete ORFs 53R and 54R (lane 2), 53R (lane 3) and 54R (lane 4) from ATV. (C.1). PCR screening of two colonies for the 53R/54R double mutant recombination cassette. A colony without the recombination cassette (lane 2) and one colony with the correctly inserted recombination cassette DNA (lane 3). (C.2). PCR colony screening of 3 colonies for the 40L recombination cassette. Each colony screened (lanes 2 – 4) contains the correct insertion of the 40L recombination cassette. LA = upstream flanking sequence; RA = downstream flanking sequence; CMV-GNR = cytomegalovirus promoter-GFP-neomycin resistance.