FIG. 4.

Optimization of annealing temperature and primer length for DD reactions. (A) Effects of different annealing temperatures on the complexity of the band pattern. RNA from toluene-induced P. putida F1 cells was processed for DD. Lanes 1 and 5, 100-bp ladder; lane 2, 30°C; lane 3, 40°C; lane 4, 50°C. (B) Effects of different primer lengths on the complexity of the band pattern. Lanes 1 and 2, 10-base primer; lanes 3 and 4, 13-base primer. RNA from uninduced cells (lanes 1 and 3) or toluene-induced cells (lanes 2 and 4) was processed for DD. The position of the todC1 fragment is indicated by arrows.