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. 2017 Apr 28;37(2):BSR20160556. doi: 10.1042/BSR20160556

Figure 2. Verification of plasmid recombination products by agarose gel separation.

Figure 2

(A) PCR amplification of hGHpA, iRFP682, MCS, CMV-promoter, and TRAIL. Lane M, DNA ladder; lane 1: PCR products of hGHPA; lane 2: PCR products of iRFP682; lane 3: PCR products of MCS; lane 4: PCR products of CMV-promoter; lane 5: PCR products of TRAIL. (B) pAAV-MCS lacking the first multiple cloning site verified by digestion. Lane 1: digested with EcoRI; lane 2: digested with SphI; lane 3: DNA ladder. (C) Plasmid lacking the second multiple cloning site verified by digestion. Lane 1: digested with BglII; lane 2: digested with BamHI; lane 3: DNA ladder. (D) Plasmid with double terminator verified by digestion. Lane 1: undigested; lane 2: digested with PstI and XhoI; lane 3: DNA ladder. (E) Plasmid with iRFP682 gene verified by digestion. Lane 1: undigested; lane 2: digested with SalI and XhoI; lane 3: DNA ladder. (F) Plasmid with new multiple cloning site verified by digestion. Lane 1: undigested; lane 2: digested with AfeI; lane 3: DNA ladder. (G) Plasmid with two complete expression cassettes verified by digestion. Lane 1: undigested; Lane 2: digested with BamHI and MluI; lane 3: DNA ladder. (H) Recombinant vector pAAV-iRFP682-TRAIL verified by digestion. Lane 1: undigested; lane 2: digested with AfeI; Lane 3: DNA ladder. (I) Recombinant vector pAAV-iRFP682-IRES-TRAIL verified by digestion. Lane 1: undigested; lane 2: digested with AscI and SphI; lane 3: DNA ladder.