npr-1(ky13) animals were mutagenized in 47 mM ethyl methanesulfonate (EMS) for 4 hours. Second generation progeny were screened for failure to develop AFT: Worms were incubated for 90 minutes in 300 mM ethanol in M9, then were placed on one side of a 10 cm Petri plate filled with assay agar [38] and 300 mM ethanol. Worms were enticed to move across the plate to a spot of chemoattractant (1 µL of 1∶200 benzaldehyde:ethanol). In these conditions, all npr-1(ky13) animals are able to develop sufficient AFT to be able to move to the spot of chemoattractant. After 90 minutes, worms that had not reached the chemoattractant were picked individually and allowed to generate self-progeny. These progeny were tested in a secondary screen for the development of AFT; animals that failed to develop AFT were kept.
