Table 2.
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) |