Figure 2

High resolution physical mapping by QDFM to linear and circular DNA molecules. A)-D) Isolated molecules of RMC11P010 were hybridized with plasmid probe pADJ762 (red) and a probe DNA specific for the P1 vector pSac BII (green). The entire P1 DNA molecules were counterstained by hybridization of random-primed RMC11P010 DNA (blue). A PCR generated ~1400 bp probe that maps in the pAd10SacBII vector near the T7 promoter site was included in the hybridization mix to show the vector orientation. A linear DNA molecule is presented in A). E) Schematic representation of the hybridization result shown in B-D). The approximate location of the Not1 site in a 'normal' P1 is indicated. F) Analysis of the overlap between two linked P1 clones ('3012', '3015') that contain one or more Not1 sites in the insert. The labeling and detection scheme is described in Table 3. G)-J) The red, green and blue images of hybridization signals along a single, closed circular DNA molecule ('3012') as well as the RGB image (J) generated by superposition of images G)-I). K) Schematic diagram showing the overlap between clones '3012' and '3015' and the location of P1-vector-specific probes.