The Selection and Use of Sorghum (Sorghum propinquum) Bacterial Artificial Chromosomes as Cytogenetic FISH Probes for Maize (Zea mays L.)

Figure 3

Southern blot confirmation of sorghum BACs after filter hybridization. Step 5 (Table 2) of the BAC selection procedure is illustrated with CBM1.11 (umc161a). (a) Sorghum propinquum FPC contig map showing contig number 147 and the five contiguous BACs (boxed) detected by filter hybridization with the CBM1.11 probe. Lane numbers are indicated beside the BACs. (b) Ethidium bromide-stained agarose gel of HindIII-digested BAC minipreps. This gel includes the BACs under investigation (lanes 9–13) along with another set (lanes 1–8) serving as negative controls. For each confirmation blot, a positive control lane [CBM1.11 (umc161a)] is included. It contains a trace amount of the same insert preparation that was used in this experiment and in the preceding filter hybridization. (c) Autoradiograph after hybridization with the CBM1.11 probe. The asterisk indicates the BAC that was eventually selected as part of step 6 (Table 2).