Fig. 7
From: A rapid in vitro method to flip back the double-floxed inverted open reading frame in a plasmid
In vitro recombination assays: Effects of DIO-flanked sequence (a) and inversion of ORF in SIO plasmids (b). a Inversion of ORF in DIO is not sequence dependent. Two plasmids (#4 and #5) with similar DIO sizes were compared. (A1) Example gel. Plasmid #4 was digested with BamHI, XhoI and BstXI. Plasmid #5 was digested with BamHI and EcoRV. (A2) Comparison of total recombination products (RT) between the two DIO plasmids. The results are averaged from three independent experiments. Error bars mean S.E.M. b Cre mediated inversion of ORF in SIO plasmids. (B1) Diagrams of SIO plasmids and Cre-mediated inversion. (B2) Comparison of the rate of ORF inversion between the two SIO plasmids. Bar chart shows the percentage of colonies with ORF inversion. Results are averaged from three independent screens. (B3) Example gel image. In vitro reactions were conducted using the standard protocol. SIO plasmids were cut with PstI. Lane 1, 3, 5 were control samples without Cre treatment. Lane 2, 4, 6 were samples with Cre treatment. The 824 bp band 740 bp band representing recombination products of two SIO plasmids are marked by R1 and R2 respectively. (B4) Bar chart of R1/R2 ratios. The results are averaged from three independent experiments. Error bars mean S.E.M. *, t-test, p < 0.05