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. 2010 Dec 22;2011:386862. doi: 10.1155/2011/386862

Table 2.

Major steps in the identification and maize marker-based selection of a sorghum BAC for use as a transgenomic FISH probe in maize. The figures and tables corresponding to the steps are indicated.

Step Procedure
1 Grow maize CBM-RFLP cultures and purify insert
(Figure 1; Table 1)
2 Radiolabel CBM insert probes for use in filter library
(step 3) or Southern blot (step 5) hybridizations
3 Hybridize probes (Tm-25°C) to Sorghum propinquum
BAC library (YRL, 36,000 BACs, ~6 × coverage;
Figure 2)
4 Score autoradiographs to identify homologous
sorghum BAC and examine location on the FPC map
(Figure 2; Table 3)
5 Grow identified overlapping BACs and verify them by
Southern blot (Figure 3)
6 Select one BAC for use as a transgenomic FISH probe
on maize pachytene chromosome spreads from maize
addition lines of oat (Figure 4)