Figure 2

Isolation of a CG15000/dNAB null allele. (a) Three differently labeled PCR reactions were pooled and concomitantly analyzed on an ABI 377 sequencer. Green, HEX-labeled white fragment (control reaction); yellow, NED-labeled CG15000 products; blue, FAM-labeled CG33273/DILP5 fragment. Color channels are not completely tight and strong products exhibit translucence. Arrows at the screenshot and the magnified inset mark a shorter product specific for one reaction. (b) One F2 fly from each cross (1 to 5) of the primary positive pool (P) was analyzed in the context of further F1 screening. Number 5 harbors the deletion allele identified in the pool. The larger, but also specific, fragment in the pool might represent a PCR artifact. The specific band marked by an asterisk turned out to be a PCR artifact as well. (c) Genomic organization of wild-type (WT) and mutant dNAB/CG15000. Exons are depicted and coding region is symbolized by filled rectangles. Small half arrows indicate primer-binding sites of the PCR-amplified region. The deleted region is filled yellow and the sequence of the wild-type and the mutant alleles starting at an ATG originally annotated as the first codon in the CG15000 ORF are aligned below.