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

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)