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. 2002 Oct 7;99(21):13498–13503. doi: 10.1073/pnas.212474399

Figure 2.

Figure 2

Screening of the extracted DNA to distinguish wtAR from floxed AR. (A) AR fragment and the flanking region. The restriction fragment of KpnI in wt is 9 kb. Three lox sites and an artificial KpnI restriction site are in the floxed AR fragment. The KpnI restriction resulted in one 7-kb and one 4-kb fragment in floxed AR. By using the 3′ end sequence as the probe (Pb), the Southern blot hybridization displayed a 9-kb fragment from ES cell clones containing wtAR, and a strong 7-kb fragment from specifically recombined ES cells containing floxed AR plus a weak 9-kb fragment from cocultured STO cells. (B) Southern blot screening of ES cells transfected with floxed AR: B3 and B9 clones are recombinated specifically and displayed a strong signal at the 7-kb position from ES cells with floxed AR and a weak signal at the 9-kb position from cocultured STO cells. B4 to B8 are the wt ES clones that displayed signal only at the 9-kb position. (C) Southern blot screening of the floxed AR clone transfected with pCMV plasmid. The pCMV-Cre restricted the sequence between two lox sites and generated four types. Lane 1 is without recombination (7 kb), lane 2 is type I recombination (5 kb), lane 3 is type II (11 kb), and lane 4 is the type III recombination (9 kb).